Secreted and transmembrane polypeptides and nucleic acids encoding the same

ABSTRACT

The present invention is directed to novel polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

FIELD OF THE INVENTION

[0001] The present invention relates generally to the identification andisolation of novel DNA and to the recombinant production of novelpolypeptides encoded by that DNA.

BACKGROUND OF THE INVENTION

[0002] Extracellular proteins play an important role in the formation,differentiation and maintenance of multicellular organisms. The fate ofmany individual cells, e.g., proliferation, migration, differentiation,or interaction with other cells, is typically governed by informationreceived from other cells and/or the immediate environment. Thisinformation is often transmitted by secreted polypeptides (for instance,mitogenic factors, survival factors, cytotoxic factors, differentiationfactors, neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Thesesecreted polypeptides or signaling molecules normally pass through thecellular secretory pathway to reach their site of action in theextracellular environment.

[0003] Secreted proteins have various industrial applications, includingpharmaceuticals, diagnostics, biosensors and bioreactors. Most proteindrugs available at present, such as thrombolytic agents, interferons,interleukins, erythropoietins, colony stimulating factors, and variousother cytokines, are secretory proteins. Their receptors, which aremembrane proteins, also have potential as therapeutic or diagnosticagents. Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. Examples of screening methods andtechniques are described in the literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)].

[0004] Membrane-bound proteins and receptors can play an important rolein the formation, differentiation and maintenance of multicellularorganisms. The fate of many individual cells, e.g., proliferation,migration, differentiation, or interaction with other cells, istypically governed by information received from other cells and/or theimmediate environment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor.

[0005] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction. Efforts are being undertaken by both industry and academiato identify new, native receptor proteins. Many efforts are focused onthe screening of mammalian recombinant DNA libraries to identify thecoding sequences for novel receptor proteins.

[0006] We herein describe the identification and characterization ofnovel secreted and transmembrane polypeptides and novel nucleic acidsencoding those polypeptides.

[0007] 1. PRO213

[0008] Human growth arrest-specific gene 6 (gas6) encodes a protein thatis expressed in a variety of different tissues and which has beenreported to be highly expressed during periods of serum starvation andnegatively regulated during growth induction. See Manfioletti et al.,Mol. Cell. Biol. 13(8):4976-4985 (1993) and Stitt et al., Cell80:661-670 (1995). Manfioletti et al. (1993), supra, have suggested thatthe gas6 protein is member of the vitamin Kependent family of proteins,wherein the members of the latter family of proteins (which include, forexample, Protein S, Protein C and Factor X) all play regulatory roles inthe blood coagulation pathway. Thus, it has been suggested that gas6 mayplay a role in the regulation of a protease cascade relevant in growthregulation or in the blood coagulation cascade. Given the physiologicalimportance of the gas6 protein, efforts are currently being undertakenby both industry and academia to identify new, native proteins which arehomologous to gas6. Many of these efforts are focused on the screeningof mammalian recombinant DNA libraries to identify the coding sequencesfor novel secreted and membrane-bound receptor proteins, specificallythose having homology to gas6. Examples of such screening methods andtechniques are described in the literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)]. We herein describe the identification of a novelpolypeptide which has homology to the gas6 polypeptide.

[0009] 2. PRO274

[0010] The 7-transmembrane (“7TM”) proteins or receptors, also referredto in the literature as G-protein coupled receptors, are specializedproteins designed for recognition of ligands and the subsequent signaltransduction of information contained within those ligands to themachinery of the cell. The primary purpose of cell surface receptors isto discriminate appropriate ligands from the various extracellularstimuli which each cell encounters, then to activate an effector systemthat produces an intracellular signal, thereby controlling cellularprocesses. [Dohlman, H., Ann. Rev. Biochem., 60:653 (1991)]. The abilityof 7TM receptors to bind ligand to a recognition domain andallosterically transmit the information to an intracellular domain is aspecialized feature of 7TM proteins [Kenakin, T., Pharmacol. Rev. 48:43(1996)]. The gene family which encodes the 7TM receptors or G-proteinlinked receptors encode receptors which recognize a large number ofligands, including but not limited to, C5a, interleukin 8 and relatedchemokines. Research in this area suggests that distinct signals at thecell surface feed into common pathways of cell activation. [Gerard, C.and Gerard, N., Curr. Op. Immunol., 6:140 (1994), Gerard, C. and Gerard,N., Ann. Rev. Immunol., 12:775 (1994)]. The superfamily of 7TM orG-protein coupled receptors contains several hundred members able torecognize various messages such as photons, ions and amino acids amongothers [Schwartz, T. W., et al., H., Trends in Pharmacol. Sci.,17(6):213 (1996)].

[0011] [Dohman, H., Ann. Rev. Biochem. 60:653 (1991)]. [Schwartz, T. W.,et al., H., Eur. J. Pharm. Sci., 2:85 (1994)]. We describe herein theidentification of a novel polypeptide (designated herein as PRO274)which has homology to the 7 transmembrane segment receptor proteins andthe Fn54 protein.

[0012] 3. PRO300

[0013] The Diff 33 protein is over-expressed in mouse testicular tumors.At present its role is unclear, however, it may play a role in cancer.Given the medical importance of understanding the physiology of cancer,efforts are currently being under taken to identify new, native proteinswhich are involved in cancer. We describe herein the identification of anovel polypeptide which has homology to Diff 33, designated herein asPRO300.

[0014] 4. PRO284

[0015] Efforts are currently being undertaken to identify andcharacterize novel transmembrane proteins. We herein describe theidentification and characterization of a novel transmembranepolypeptide, designated herein as PRO284.

[0016] 5. PRO296

[0017] Cancerous cells often express numerous proteins that are notexpressed in the corresponding normal cell type or are expressed atdifferent levels than in the corresponding normal cell type. Many ofthese proteins are involved in inducing the transformation from a normalcell to a cancerous cell or in maintaining the cancer phenotype. Assuch, there is significant interest in identifying and characterizingproteins that are expressed in cancerous cells. We herein describe theidentification and characterization of a novel polypeptide havinghomology to the sarcoma-amplified protein SAS, designated herein asPRO296.

[0018] 6. PRO329

[0019] Immunoglobulin molecules play roles in many important mammalianphysiological processes. The structure of immunoglobulin molecules hasbeen extensively studied and it has been well documented that intactimmunoglobulins possess distinct domains, one of which is the constantdomain or F_(c) region of the immunoglobulin molecule. The F_(c) domainof an immunoglobulin, while not being directly involved in antigenrecognition and binding, does mediate the ability of the immunoglobulinmolecule, either uncomplexed or complexed with its respective antigen,to bind to F_(c) receptors either circulating in the serum or on thesurface of cells. The ability of an F_(c) domain of an immunoglobulin tobind to an F_(c) receptor molecule results in a variety of importantactivities, including for example, in mounting an immune responseagainst unwanted foreign particles. As such, there is substantialinterest in identifying novel F_(c) receptor proteins and subunitsthereof. We herein describe the identification and characterization of anovel polypeptide having homology to a high affinity immunoglobulinF_(c) receptor protein, designated herein as PRO329.

[0020] 7. PRO362

[0021] Colorectal carcinoma is a malignant neoplastic disease which hasa high incidence in the Western world, particularly in the UnitedStates. Tumors of this type often metastasize through lymphatic andvascular channels and result in the death of some 62,000 persons in theUnited States annually.

[0022] Monoclonal antibody A33 (mAbA33) is a murine immunoglobulin thathas undergone extensive preclinical analysis and localization studies inpatients inflicted with colorectal carcinoma (Welt et al., J. Clin.Oncol. 8:1894-1906 (1990) and Welt et al., J. Clin. Oncol. 12:1561-1571(1994)). mAbA33 has been shown to bind to an antigen found in and on thesurface of normal colon cells and colon cancer cells. In carcinomasoriginating from the colonic mucosa, the A33 antigen is expressedhomogeneously in more than 95% of the cases. The A33 antigen, however,has not been detecting in a wide range of other normal tissues, i.e.,its expression appears to be rather organ specific. Therefore, the A33antigen appears to play an important role in the induction of colorectalcancer.

[0023] Given the obvious importance of the A33 antigen in tumor cellformation and/or proliferation, there is substantial interest inidentifying homologs of the A33 antigen. In this regard, we hereindescribe the identification and characterization of a novel polypeptidehaving homology to the A33 antigen protein, designated herein as PRO362.

[0024] 8. PRO363

[0025] The cell surface protein HCAR is a membrane-bound protein thatacts as a receptor for subgroup C of the adenoviruses and subgroup B ofthe coxsackieviruses. Thus, HCAR may provide a means for mediating viralinfection of cells in that the presence of the HCAR receptor on thecellular surface provides a binding site for viral particles, therebyfacilitating viral infection.

[0026] In light of the physiological importance of membrane-boundproteins and specifically those which serve a cell surface receptor forviruses, efforts are currently being undertaken by both industry andacademia to identify new, native membrane-bound recptor proteins. Manyof these efforts are focused on the screening of mammalian recombinantDNA libraries to identify the coding sequences for novel receptorproteins. We herein describe a novel membrane-bound polypeptide havinghomology to the cell surface protein HCAR and to various tumor antigensincluding A33 and carcinoembryonic antigen, designated herein as PRO363,wherein this polypeptide may be a novel cell surface virus receptor ortumor antigen.

[0027] 9. PRO868

[0028] Control of cell numbers in mammals is believed to be determined,in part, by a balance between cell proliferation and cell death. Oneform of cell death, sometimes referred to as necrotic cell death, istypically characterized as a pathologic form of cell death resultingfrom some trauma or cellular injury. In contrast, there is another,“physiologic” form of cell death which usually proceeds in an orderly orcontrolled manner. This orderly or controlled form of cell death isoften referred to as “apoptosis” [see, e.g., Barr et al.,Bio/Technology, 12:487-493 (1994); Steller et al., Science,267:1445-1449 (1995)]. Apoptotic cell death naturally occurs in manyphysiological processes, including embryonic development and clonalselection in the immune system [Itoh et al., Cell, 66:233-243 (1991)].Decreased levels of apoptotic cell death have been associated with avariety of pathological conditions, including cancer, lupus, and herpesvirus infection [Thompson, Science, 267:1456-1462 (1995)]. Increasedlevels of apoptotic cell death may be associated with a variety of otherpathological conditions, including AIDS, Alzheimer's disease,Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis,retinitis pigmentosa, cerebellar degeneration, a plastic anemia,myocardial infarction, stroke, reperfusion injury, and toxin-inducedliver disease [see, Thompson, supra].

[0029] Apoptotic cell death is typically accompanied by one or morecharacteristic morphological and biochemical changes in cells, such ascondensation of cytoplasm, loss of plasma membrane microvilli,segmentation of the nucleus, degradation of chromosomal DNA or loss ofmitochondrial function. A variety of extrinsic and intrinsic signals arebelieved to trigger or induce such morphological and biochemicalcellular changes [Raff, Nature, 356-397-400 (1992); Steller, supra;Sachs et al., Blood, 82:15 (1993)]. For instance, they can be triggeredby hormonal stimuli, such as glucocorticoid hormones for immaturethymocytes, as well as withdrawal of certain growth factors[Watanabe-Fukunaga et al., Nature 356:314-317 (1992)]. Also, someidentified oncogenes such as Myc, rel, and E1A, and tumor suppressors,like p53, have been reported to have a role in inducing apoptosis.Certain chemotherapy drugs and some forms of radiation have likewisebeen observed to have apoptosis-inducing activity [Thompson, supra].

[0030] Various molecules, such as tumor necrosis factor-α (“TNF-α”),tumor necrosis factory (“TNF-β” or “lymphotoxin-α”), lymphotoxin-β(“LT-β”), CD30 ligand, CD27 ligand, CD40 ligand, OX-40 ligand, 4-1BBligand, Apo-1 ligand (also referred to as Fas ligand or CD95 ligand),and Apo-2 ligand (also referred to as TRAIL) have been identified asmembers of the tumor necrosis factor (“TNF”) family of cytolines [See,e.g., Gruss and Dower, Blood, 85:3378-3404 (1995); Pitti et al., J.Biol. Chem., 271:12687-12690 (1996); Wiley et al., Immunity, 3.673-682(1995); Browning et al., Cell, 72:847-856 (1993); Armitage et al.Nature, 357:80-82 (1992), WO 97101633 published Jan. 16, 1997; WO97/25428 published Jul. 17, 19971. Among these molecules, TNF-α, TNF-β,CD30 ligand, E1BB ligand, Apo-1 ligand, and Apo-2 ligand (TRAEL) havebeen reported to be involved in apoptotic cell death. Both TNF-α andTNF-β have been reported to induce apoptotic death in susceptible tumorcells [Schmid et al., Proc. Natl. Acad. Sci., 83:1881 (1986); Dealtry etal., Eur. J. Immunol., 17:689 (1987)]. Zheng et al. have reported thatTNF-α is involved in post-stimulation apoptosis of CD8-positive T cells[Zheng et al., Nature, 377:348-351 (1995)]. Other investigators havereported that CD30 ligand may be involved in deletion of self-reactive Tcells in the thymus [Amakawa et al., Cold Spring Harbor LaboratorySymposium on Programmed Cell Death, Abstr. No. 10, (1995)].

[0031] Mutations in the mouse Fas/Apo-1 receptor or ligand genes (calledIpr and gld, respectively) have been associated with some autoimmunedisorders, indicating that Apo-1 ligand may play a role in regulatingthe clonal deletion of self-reactive lymphocytes in the periphery rammeret al., Curr. Op. Immunol., 6:279-289 (1994); Nagata et al., Science,267:1449-1456 (1995)]. Apo-1 ligand is also reported to inducepost-stimulation apoptosis in CD4-positive T lymphocytes and in Blymphocytes, and may be involved in the elimination of activatedlymphocytes when their function is no longer needed [Krammer et al.,supra; Nagata et al., supra]. Agonist mouse monoclonal antibodiesspecifically binding to the Apo-1 receptor have been reported to exhibitcell killing activity that is comparable to or similar to that of TNF-α[Yonehara et al., J. Exp. Med., 169:1747-1756 (1989)].

[0032] Induction of various cellular responses mediated by such TNFfamily cytokines is believed to be initiated by their binding tospecific cell receptors. Two distinct TNF receptors of approximately55-kDa (TNFR1) and 75-kDa (TNFR2) have been identified [Holman et al.,J. Biol. Chem., 264:14927-14934 (1989); Brockhaus et al., Proc. Natl.Acad. Sci., 87:3127-3131 (1990); EP 417,563, published Mar. 20, 19911and human and mouse cDNAs corresponding to both receptor types have beenisolated and characterized [Loetscher et al., Cell, 61:351 (1990);Schall et al., Cell, 61:361 (1990); Smith et al., Science, 248:1019-1023(1990); Lewis et al., Proc. Natl. Acad. Sci., 88:2830-2834 (1991);Goodwin et al., Mol. Cell. Biol., 11:3020-3026 (1991)]. Extensivepolymorphisms have been associated with both TNF receptor genes [see,e.g., Takao et al., Immunogenetics, 37:199-203 (1993)]. Both TNFRs sharethe typical structure of cell surface receptors including extracellular,transmembrane and intracellular regions. The extracellular portions ofboth receptors are found naturally also as soluble TNF-binding proteins[Nophar, Y. et al., EMBO J., 9:3269 (1990); and Kohno, T. et al., Proc.Natl. Acad. Sci. U.S.A., 87:8331 (1990)]. More recently, the cloning ofrecombinant soluble TNF receptors was reported by Hale et al. [J. Cell.Biochem. Supplement 15SF, 1991, p. 113 (P424)].

[0033] The extracellular portion of type 1 and type 2 TNFRs (TNFR1 andTNFR2) contains a repetitive amino acid sequence pattern of fourcysteine-rich domains (CRDs) designated 1 through 4, staring from theNH₂-terminus. Each CRD is about 40 amino acids long and contains 4 to 6cysteine residues at positions which are well conserved [Schall et al.,supra; Loetscher et al., supra; Smith et al., supra; Nophar et al.,supra; Kohno et al., supra . In TNFR1, the approximate boundaries of thefour CRDs are as follows: CRD1-amino acids 14 to about 53; CRD2-aminoacids from about 54 to about 97; CRD3-amino acids from about 98 to about138; CRD4-amino acids from about 139 to about 167. In TNFR2, CRD1includes amino acids 17 to about 54; CRD2-amino acids from about 55 toabout 97; CRD3-amino acids from about 98 to about 140; and CRD4-aminoacids from about 141 to about 179 [Banner et al., Cell, 73:431-435(1993)]. The potential role of the CRDs in ligand binding is alsodescribed by Banner et al., supra.

[0034] A similar repetitive pattern of CRDs exists in several othercell-surface proteins, including the p75 nerve growth factor receptor(NGFR) [Johnson et al., Cell, 47:545 (1986); Radeke et al., Nature,325:593 (1987)], the B cell antigen CD40 [Stamenkovic et al., EMBO J.,8:1403 (1989)], the T cell antigen OX40 Mallet et al., EMBO J., 9:1063(1990)] and the Fas antigen [Yonehara et al., supra and Itoh et al.,Cell, 66:233-243 (1991)]. CRDs are also found in the soluble TNFR(sTNFR)-like T2 proteins of the Shope and myxoma poxviruses [Upton etal., Virology, 160:20-29 (1987); Smith et al., Biochem. Biophys. Res.Commun., 176:335 (1991); Upton et al., Virology, 184:370 (1991)].Optimal alignment of these sequences indicates that the positions of thecysteine residues are well conserved. These receptors are sometimescollectively referred to as members of the TNF/NGF receptor superfamily.Recent studies on p75NGFR showed that the deletion of CRD1 [Welcher, A.A. et al., Proc. Natl. Acad. Sci. USA, 88:159-163 (1991)] or a 5-aminoacid insertion in this domain [Yan, H. and Chao, M. V., J. Biol. Chem.,266:12099-12104 (1991)] had little or no effect on NGF binding [Yan, H.and Chao, M. V., supra]. p75 NGFR contains a proline-rich stretch ofabout 60 amino acids, between its CRD4 and transmembrane region, whichis not involved in NGF binding [Peetre, C. et al., Eur. J. Hematol.,41:414-419 (1988); Seckinger, P. et al., J. Biol. Chem., 264:11966-11973(1989); Yan, H. and Chao, M. V., supra]. A similar proline-rich regionis found in TNFR2 but not in TNFR1.

[0035] The TNF family ligands identified to date, with the exception oflymphotoxin-α, are type II transmembrane proteins, whose C-terminus isextracellular. In contrast, most receptors in the TNF receptor (TNFR)family identified to date are type I transmembrane proteins. In both theTNF ligand and receptor families, however, homology identified betweenfamily members has been found mainly in the extracellular domain(“ECD”). Several of the TNF family cytolines, including TNF-α, Apo-1ligand and CD40 ligand, are cleaved proteolytically at the cell surface;the resulting protein in each case typically forms a homotrimericmolecule that functions as a soluble cytokine. TNF receptor familyproteins are also usually cleaved proteolytically to release solublereceptor ECDs that can function as inhibitors of the cognate cytokines.

[0036] Recently, other members of the TNFR family have been identified.Such newly identified members of the TNFR family include CAR1, HVEM andosteoprotegerin (OPG) [Brojatsch et al., Cell, 87:845-855 (1996);Montgomery et al., Cell, 87:427-436 (1996); Marsters et al., J. Biol.Chem., 272:14029-14032 (1997); Simonet et al., Cell, 89:309-319 (1997)].Unlike other known TNFR-like molecules, Simonet et al., supra, reportthat OPG contains no hydrophobic trausmembrane-spanning sequence.

[0037] Moreover, a new member of the TNFINGF receptor family has beenidentified in mouse, a receptor referred to as “GITR” for“glucocorticoid-induced tumor necrosis factor receptor family-relatedgene” [Nocentini et al., Proc. Natl. Acad. Sci. USA 94:6216-6221(1997)]. The mouse GITR receptor is a 228 amino acid type Itransmembrane protein that is expressed in normal mouse T lymphocytesfrom thymus, spleen and lymph nodes. Expression of the mouse GITRreceptor was induced in T lymphocytes upon activation with anti-CD3antibodies, Con A or phorbol 12-myristate 13-acetate. It was speculatedby the authors that the mouse GITR receptor was involved in theregulation of T cell receptor-mediated cell death.

[0038] In Marsters et al., Curr. Biol., 6:750 (1996), investigatorsdescribe a full length native sequence human polypeptide, called Apo-3,which exhibits similarity to the TNFR family in its extracellularcysteine-rich repeats and resembles TNFR1 and CD95 in that it contains acytoplasmic death domain sequence [see also Marsters et al., Curr.Biol., 6:1669 (1996)]. Apo-3 has also been referred to by otherinvestigators as DR3, ws1-1 and TRAMP [Chinnaiyan et al., Science,274:990 (1996); Kitson et al., Nature, 384:372 (1996); Bodmer et al.,Immunity, 6:79 (1997)].

[0039] Pan et al. have disclosed another TNF receptor family memberreferred to as “DR4” [Pan et al., Science, 276:111-113 (1997)]. The DR4was reported to contain a cytoplasmic death domain capable of engagingthe cell suicide apparatus. Pan et al. disclose that DR4 is believed tobe a receptor for the ligand known as Apo-2 ligand or TRAIL.

[0040] In Sheridan et al., Science, 277:818-821 (1997) and Pan et al.,Science, 277:815-818 (1997), another molecule believed to be a receptorfor the Apo-2 ligand (TRAIL) is described. That molecule is referred toas DR5 (it has also been alternatively referred to as Apo-2). Like DR4,DR5 is reported to contain a cytoplasmic death domain and be capable ofsignaling apoptosis.

[0041] In Sheridan et al., supra, a receptor called DcR1 (oralternatively, Apo-2DcR) is disclosed as being a potential decoyreceptor for Apo-2 ligand (TRAIL). Sheridan et al. report that DcR1 caninhibit Apo-2 ligand function in vitro. See also, Pan et al., supra, fordisclosure on the decoy receptor referred to as TRID.

[0042] For a review of the TNF family of cytokines and their receptors,see Gruss and Dower, supra.

[0043] As presently understood, the cell death program contains at leastthree important elements—activators, inhibitors, and effectors; in C.elegans, these elements are encoded respectively by three genes, Ced4,Ced-9 and Ced-3 [Steller, Science, 267:1445 (1995); Chinnaiyan et al.,Science, 275:1122-1126 (1997); Wang et al., Cell, 90:1-20 (1997)]. Twoof the TNFR family members, TNFR1 and Fas/Apo1 (CD95), can activateapoptotic cell death [Chinnaiyan and Dixit, Current Biology, 6:555-562(1996); Fraser and Evan, Cell; 85:781-784 (1996)]. TNFR1 is also knownto mediate activation of the transcription factor, NF-κB [Tartaglia etal., Cell, 74:845-853 (1993); Hsu et al., Cell, 84:299-308 (1996)]. Inaddition to some ECD homology, these two receptors share homology intheir intracellular domain (ICD) in an oligomerization interface knownas the death domain [Tartaglia et al., supra; Nagata, Cell, 88:355(1997)]. Death domains are also found in several metazoan proteins thatregulate apoptosis, namely, the Drosophila protein, Reaper, and themammalian proteins referred to as FADD/MORT1, TRADD, and RIP [Cleavelandand Ihle, Cell, 81:479-482 (1995)].

[0044] Upon ligand binding and receptor clustering, TNFR1 and CD95 arebelieved to recruit FADD into a death-inducing signalling complex. CD95purportedly binds FADD directly, while TNFR1 binds FADD indirectly viaTRADD [Chinnaiyan et al., Cell, 81:505-512 (1995); Boldin et al., J.Biol. Chem., 270:387-391 (1995); Hsu et al., supra; Chinnaiyan et al.,J. Biol. Chem., 271:4961-4965 (1996)]. It has been reported that FADDserves as an adaptor protein which recruits the Ced-3-related protease,MACHα/FLICE (caspase 8), into the death signalling complex [Boldin etal., Cell, 85:803-815 (1996); Muzio et al., Cell, 85:817-827 (1996)].MACHα/FLICE appears to be the trigger that sets off a cascade ofapoptotic proteases, including the interleulin-1β converting enzyme(ICE) and CPP32/Yama, which may execute some critical aspects of thecell death programme [Fraser and Evan, supra].

[0045] It was recently disclosed that programmed cell death involves theactivity of members of a family of cysteine proteases related to the C.elegans cell death gene, ced-3, and to the mammalian IL-1-convertingenzyme, ICE. The activity of the ICE and CPP32/Yama proteases can beinhibited by the product of the cowpox virus gene, cmzA [Ray et al.,Cell, 69:597-604 (1992); Tewari et al., Cell, 81:801-809 (1995)]. Recentstudies show that CrmA can inhibit TNFR1- and CD95-induced cell death[Enari et al., Nature, 375:78-81 (1995); Tewari et al., J. Biol. Chem.,270:3255-3260 (1995)].

[0046] As reviewed recently by Tewari et al., TNFR1, TNFR2 and CD40modulate the expression of proinflammatory and costimulatory cytokines,cytokine receptors, and cell adhesion molecules through activation ofthe transcription factor, NF-κB [Tewari et al., Curr. Op. Genet.Develop., 6:39-44 (1996)]. NF-κB is the prototype of a family of dimerictranscription factors whose subunits contain conserved Rel regions[Verma et al., Genes Develop., 2:2723-2735 (1996); Baldwin, Ann. Rev.Immunol., 14:649-681 (1996)]. In its latent form, NF-κB is complexedwith members of the IκB inhibitor family; upon inactivation of the IκBin response to certain stimuli, released NF-κB translocates to thenucleus where it binds to specific DNA sequences and activates genetranscription.

[0047] 10. PRO382

[0048] Proteases are enzymatic proteins which are involved in a largenumber of very important biological processes in mammalian andnon-mammalian organisms. Numerous different protease enzymes from avariety of different mammalian and non-mammalian organisms have beenboth identified and characterized, including the serine proteases whichexhibit specific activity toward various serine-containing proteins. Themammalian protease enzymes play important roles in biological processessuch-as, for example, protein digestion, activation, inactivation, ormodulation of peptide hormone activity, and alteration of the physicalproperties of proteins and enzymes.

[0049] In light of the important physiological roles played by proteaseenzymes, efforts are currently being undertaken by both industry andacademia to identify new, native protease homologs. Many of theseefforts are focused on the screening of mammalian recombinant DNAlibraries to identify the coding sequences for novel membrane-boundreceptor proteins. Examples of screening methods and techniques aredescribed in the literature [see, for example, Klein et al., Proc. Natl.Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No. 5,536,637)]. We hereindescribe the identification of novel polypeptides having homology toserine protease enzymes, designated herein as PRO382 polypeptides.

[0050] 11. PRO545

[0051] The ADAM (A Disintegrin And Metalloprotease) family of proteinsof which meltrin is a member may have an important role in cellinteractions and in modulating cellular responses. [see, for example,Gilpin et al., J. Biol. Chem., 273(1:157-166(1998)]. The ADAM proteinshave been implicated incarcinogenesis. Meltrin-α(ADAM12) is a myoblastgene product reported to be required for cell fusion. [Harris et al., J.Cell. Biochem., 67(1):136-142 (1997), Yagami-Hiromasa et al., Nature,377:652-656 (1995)]. The peltrins contain disintegrin and metaloproteasedomains and are implicated in cell adhesive events involved indevelopment, through the integrin-binding disintegrin domain, but alsohave an anti-adhesive function through a zinc-dependent metaroproteasedomain. [Alfandari et al., Devel. Biol., 1.82(2):314-330 (1997)]. Giventhe medical importance of cell fusion and modulation of cellularresponses in carcinogenesis and other disease mechanisms, efforts arecurrently being under taken to identify new, native proteins which areinvolved in cell fusion and modulation of cellular responses. Wedescribe herein the identification of a novel polypeptide which hashomology to meltrin, designated herein as PRO545.

[0052] 12. PRO617

[0053] CD24 is a protein that is associated with the cell surface of avariety of different cells of the mammalian immune system, including forexample, neutrophils, monocytes and some lymphocytes, for example, Blymphocytes. CD24 has been shown to be a ligand for theplatelet-associated surface glycoprotein P-selectin (also known asgranule membrane protein-140 or GMP-140), a glycoprotein that isconstitutively synthesized in both platelets and endothelial cells andbecomes exposed on the surface of platelets when those cells becomeactivated. In this way, P-selectin mediates the calcium-dependentadhesion of activated platelets and endothelial cells to the variouscells of the immune system that express one or more ligands for theP-selectin molecule, particularly CD24. This mechanism allows forrecruitment of immune system cells to locations where they are mostneeded, for example, sites of injury. Thus, there is substantialinterest in identifying novel polypeptides that exhibit homology to thecell surface antigens of the immune system cells. We herein describe theidentification and characterization of a novel polypeptide havinghomology to the CD24 protein, wherein that novel polypeptide is hereindesignated PRO617.

[0054] 13. PRO700

[0055] Protein-disulfide isomerase (PDI) is a catalyst of disulfideformation and isomerization during protein folding. It has two catalyticsites housed in two domains homologous to thioredoxin, one near the Nterminus and the other near the C terminus. [See for example, GilbertHF, J.Biol.Chem., 47:29399-29402 (1997), Mayfield K J, Science,278:1954-1957 (1997) and Puig et al., J.Biol.Chem., 52:32988-32994(1997)]. PDI is useful for formation of natural type disulfide bonds ina protein which is produced in aprokaryotic cell. (See also, U.S. Pat.Nos. 5,700,659 and 5,700,678).

[0056] Thus, PDI and molecules related thereto are of interest,particularly for ability to catalyze the formation of disulfide bonds.Moreover, these molecules are generally of interest in the study ofredox reactions and related processes. PDI and related molecules arefurther described in Darby, et al., Biochemistry 34, 11725-11735(1995).We herein describe the identification and characterization of novelpolypeptides having homology to protein disulfide isomerase, designatedherein as PRO700 polypeptides.

[0057] 14. PRO702

[0058] Conglutinin is a bovine serum protein that was originallydescribed as a vertebrate lectin protein and which belongs to the familyof C-type lectins that have four characteristic domains, (1) anN-terminal cysteine-rich domain, (2) a collagen-like domain, (3) a neckdomain and (4) a carbohydrate recognition domain (CRD). Recent reportshave demonstrated that bovine conglutinin can inhibit hemagglutinationby influenza A viruses as a result of their lectin properties (Eda etal., Biochem. J. 316:4348 (1996)). It has also been suggested thatlectins such as conglutinin can function as immunoglobulin-independentdefense molecules due to complement- mediated mechanisms. Thus,conglutinin has been shown to be useful for purifying immune complexesin vitro and for removing circulating immune complexes from patientsplasma in vivo (Lim et al., Biochem. Biophys. Res. Commun. 218:260-266(1996)). We herein describe the identification and characterization of anovel polypeptide having homology to the conglutinin protein, designatedherein as PRO702.

[0059] 15. PRO703

[0060] Very-long-chain acyl-CoA synthetase (“VLCAS”) is a long-chainfatty acid transport protein which is active in the cellular transportof long and very long chain fatty acids. [see for example, Uchida etal., J. Biochem (Tokyo) 119(3):565-571 (1996) and Uchiyama et al., JBiol Chem 271(48):30360-30365 (1996). Given the biological importance offatty acid transport mechanisms, efforts are currently being under takento identify new, native proteins which are involved in fatty acidtransport. We describe herein the identification of a novel polypeptidewhich has homology to VLCAS, designated herein as PRO703.

[0061] 16. PRO705

[0062] The glypicans are a family of glycosylphosphatidylinositol(GPI)-anchored proteoglycans that, by virtue of their cell surfacelocalization and possession of heparin sulfate chains, may regulate theresponses of cells to numerous heparin-binding growth factors, celladhesion molecules and extracellular matrix components. Mutations in oneglypican protein cause of syndrome of human birth defects, suggestingthat the glypicans may play an important role in development (Litwack etal., Dev. Dyn. 211:72-87 (1998)). Also, since the glypicans may interactwith the various extracellular matrices, they may also play importantroles in wound healing (McGrath et al., Pathol. 183:251-252 (1997)).Furthermore, since glypicans are expressed in neurons and glioma cells,they may also play an important role in the regulation of cell divisionand survival of cells of the nervous system (Liang et al., J. Cell.Biol. 139:851-864 (1997)). It is evident, therefore, that the glypicansare an extremely important family of proteoglycans. There is, therefore,substantial interest in identifying novel polypeptides having homologyto members of the glypican family. We herein describe the identificationand characterization of a novel polypeptide having homology toK-glypican, designated herein as PRO705.

[0063] 17. PRO708

[0064] Aryl sulfatases are enzymes that exist in a number of differentisoforms, including aryl sulfatase A (ASA), aryl sulfatase B (ASB) andaryl sulfatase C (ASC), and that function to hydrolyze a variety ofdifferent aromatic sulfates. Aryl sulfatases have been isolated from avariety of different animal tissues and microbial sources and theirstructures and functions have been extensively studied (see, e.g.,Nichol and Roy, J. Biochem. 55:643-651 (1964)). ASA deficiency has beenreported to be associated with metachromatic leukodystrophy (MLD) (Gileset al., Prenat. Diagn. 7(4):245-252 (1987) and Herska et al., Am. J.Med. Genet. 26(3):629-635 (1987)). Additionally, other groups havereported that aryl sulfatases have been found in high levels in naturalkiller cells of the immune system and have hypothesized a possible rolefor these enzymes in NK cell-mediated cellular lysis (see, e.g.,Zucker-Franklin et al., Proc. Natl. Acad. Sci. USA 80(22):6977-6981(1983)). Given the obvious physiological importance of the arylsulfatase enzymes, there is a substantial interest in identifying novelaryl sulfatase homolog polypeptides. We herein describe theidentification and characterization of novel polypeptides havinghomology to the aryl sulfatases, wherein these novel polypeptides areherein designated PRO708 polypeptides.

[0065] 18. PRO320

[0066] Fibulin-1 is a cysteine-rich, calcium-binding extracellularmatrix (ECM) component of basement membranes and connective tissueelastic fibers and plasma protein, which has four isoforms, A-D, derivedfrom alternative splicing. Fibulin-1 is a modular glycoprotein withamino-terminal anaphlatoxin-like modules followed by nine epidermalgrowth factor (EGF)-like modules and, depending on alternative splicing,four possible carboxyl termini. Fibulin-2 is a novel extracellularmatrix protein frequently found in close association with microfibrilscontaining either fibronectin or fibrillin. There are multiple forms offibulin-1 that differ in their C-terminal regions that are producedthrough the process of alternative splicing of their precursor RNA. [seefor example Tran et al., Matrix Biol 15(7):479-493 (1997).]

[0067] Northern and Western blotting analysis of 16 cell linesestablished from tumors formed in athymic mice and malignant cell linesderived from patients indicate that low expression of fibulin-1D plays arole in tumor formation and invasion. [Qing et al., Oncogene,18:2159-2168 (1997)]. Ovarian-cancer cells are characterized by theirability to invade freely the peritoneal cavity. It has been demonstratedthat estradiol stimulates the proliferation of estrogen-receptor(ER)-positive ovarian-cancer cells, as well as expression of fibulin-1.Studies on the effect of fibulin-1 on motility of the MDA-MB231breast-cancer cell line, indicated inhibition of haptotactic migrationof MDA-MB231 cells, and the authors concluded that fibulin-1 can inhibitcancer cell motility in vitro and therefore has the potential to inhibittumor invasion. [Hayashido et al., Int J Cancer, 75(4):654-658 (1998)]

[0068] Thus, fibulin, and molecules related thereto are of interest,particularly for the use of preventing cancer. Moreover, these moleculesare generally of interest in the study of connective tissue andattachment molecules and related mechanisms. Fibulin and relatedmolecules are further described in Adams, et al., J. Mol. Biol.,272(2):226-36 (1997); Kielty and Shuttleworth, Microse. Res. Tech.,38(4):413-27 (1997); and Child. J. Card. Surg. 12(2Supp.):131-5 (1997).

[0069] We herein describe the identification and characterization ofnovel polypeptides having homology to fibulin, designated herein asPRO320 polypeptides.

[0070] 19. PRO324

[0071] Oxidoreductases are enzymes that catalyze a reaction in which twomolecules of a compound interact so that one molecule is oxidized andthe other is reduced, with a molecule of water entering the reaction.There are many different types of oxidoreductase enzymes that play veryimportant physiological roles in the mammalian organism. Some of themost important oxidoreductases include, for example, lyases, lactases,cholesterol oxidases, and the like. These enzymes play roles in suchessential processes as digestion, signal transduction, maintenance ofionic homeostasis, and the like. As such, given that oxidoreductaseenzymes find various essential uses in the mammalian organism, there isa substantial interest in identifying novel oxidoreductase enzymehomologs. We herein describe the identification and characterization ofa novel polypeptide having homology to oxidoreductases, designatedherein as PRO324.

[0072] 20. PRO351

[0073] Prostasin is a novel human serine proteinase purified from humanseminal fluid. Immunohistochemical localization reveals that prostasinis present in epithelial cells and ducts of the prostate gland. The cDNAfor prostasin has been cloned and characterized. Southern blot analysis,following a reverse transcription polymerase chain reaction, indicatesthat prostasin mRNA is expressed in prostate, liver, salivary gland,kidney, lung, pancreas, colon, bronchus, renal proximal tubular cells,and prostate carcinoma LNCαP cells. Cellular localization of prostasinmRNA was identified within epithelial cells of the human prostate glandby in situ hybridization histochemistry. [See for example, Yu et al., JBiol Chem. (1994) 269(29):18843-18848, and Yu et al., J Biol Chem.(1994) 270(22):13483-13489].

[0074] Thus, prostasin, and molecules related thereto are of interest,particularly for the study, diagnosis and treatment of medicalconditions involving the prostate. Prostasin and related molecules arefurther described in Yu et al., Genomics (1996) 32(3):334-340. We hereindescribe the identification and characterization of novel polypeptideshaving homology to prostasin, designated herein as PRO351 polypeptides.

[0075] 21. PRO352

[0076] Butyrophilin is a milk glycoprotein that constitutes more than40% of the total protein associated with the fat globule membrane inmammalian milk. Expression of butyrophilin mRNA has been shown tocorrelate with the onset of milk fat production toward the end pregnancyand is maintained throughout lactation. Butyrophilin has been identifiedin bovine, murine and human (see Taylor et al., Biochim. Biophys. Acta1306:14 (1996), Ishii et al., Biochim. Biophys. Acta 1245:285-292(1995), Mather et al., J. Dairy Sci. 76:3832-3850 (1993) and Banghart etal., J. Biol. Chem. 273:4171-4179 (1998)) and is a type I transmembraneprotein that is incorporated into the fat globulin membrane. It has beensuggested that butyrophilin may play a role as the principle scaffoldfor the assembly of a complex with xanthine dehydrogenase/oxidase andother proteins that function in the budding and release of milk-fatglobules from the apical surface during lactation (Banghart et al.,supra .

[0077] Given that butyrophilin plays an obviously important role inmammalian milk production, there is substantial interest in identifyingnovel butyrophilin homologs. We herein describe the identification andcharacterization of a novel polypeptide having homology to butyrophilin,designated herein as PRO352.

[0078] 22. PRO381

[0079] The immunophilins are a family of proteins that function asreceptors for immunosuppressant drugs, such as cyclosporin A, FK506, andrapamycin. The immunophilins occur in two separate classes, (1) theFK506binding proteins (FKBPs), which bind to FK506 and rapamycin, and(2) the cyclophilins, which bind to cyclosporin A. With regard to theFK506-binding proteins, it has been reported that the FK506/FKBP complexfunctions to inhibit the activity of the serine/threonine proteinphosphatase 2B (calcineurin), thereby providing immunosuppressantactivity (Gold, Mol. Neurobiol. 15:285-306 (1997)). It has also beenreported that the FKBP immunophilins are found in the mammalian nervoussystem and may be involved in axonal regeneration in the central nervoussystem through a mechanism that is independent of the process by whichimmunosuppression is achieved (Gold, supra). Thus, there is substantalinterest in identifying novel polypeptides having homology to the FKBPimmunophilins. We herein describe the identification andcharacterization of a novel polypeptide having homology to an FKBPimmunophilin protein, designated herein as PRO381.

[0080] 23. PRO386

[0081] Mammalian cell membranes perform very important functionsrelating to the structural integrity and activity of various cells andtissues. Of particular interest in membrane physiology is the study oftransmembrane ion channels which act to directly control a variety ofphysiological, pharmacological and cellular processes. Numerous ionchannels have been identified including calcium (Ca), sodium (Na) andpotassium (K) channels, each of which have been analyzed in detail todetermine their roles in physiological processes in vertebrate andinsect cells.

[0082] One type of cell membrane-associated ion channel, the sodiumchannel, plays an extremely important role in a cell's ability tomaintain ionic homeostasis as well as transmit intracellular andextracellular signals. Voltage-gated sodium channels in brain neuronshave been shown to be complexes of a pore-forming alpha unit withsmaller beta-1 and beta-2 subunits (Isom et al., Cell 83:433-442(1995)). Given the obvious importance of sodium channels in cellularhomeostasis and other important physiological functions, there is asignificant interest in identifying novel polypeptides having homologyto sodium channel subunits. We herein describe the identification andcharacterization of a novel polypeptide having homology to the beta-2subunit of the rat sodium channel, designated herein as PRO386.

[0083] 24. PRO540

[0084] Lecithin-cholesterol acyltransferase (“LCAT”), also known asphosphatidylcholine-sterol acyltransferase is a key enzyme in theintravascular metabolism of high density lipoproteins, specifically inthe process of cholesterol metabolism. [see, for example, Brousseau etal., J. Lipid Res., 38(12):2537-2547 (1997), Hill et al., Biochem. J.,294:879-884 (1993), and Drayna et al., Nature 327 (6123):632-634(1987)]. Given the medical importance of lipid metabolism, efforts arecurrently being under taken to identify new, native proteins which areinvolved in lipid transport. We describe herein the identification of anovel polypeptide which has homology to LCAT, designated herein asPRO540.

[0085] 25. PRO615

[0086] Synaptogyrin is a synaptic vesicle protein that is uniformlydistributed in the nervous system. The cDNA encoding synaptogyrin hasbeen cloned and sequenced and the sequence predicts a protein with amolecular mass of 25,900 D with four membrane-spanning domains.Synaptogyrin has been implicated in membrane traffic to and from theplasma membrane. Stenius et al., J. Cell. Biol. 131(6-2):1801-1809(1995). In addition, a novel isoform of synaptogyrin called cellugyrinexhibits sequence identity with synaptogyrin. In rat tissues, cellugyrinand synaptogyrins are expressed in mirror image patterns. Cellugyrin isubiquitously present in all tissues tested with the lowest levels inbrain tissue, whereas synaptogyrin protein is only detectable in brain.In rat tissues, cellugyrin and synaptogyrins are expressed in mirrorimage patterns. The synaptic vesicle protein synaptogyrin may be aspecialized version of a ubiquitous protein, cellugyrin, with the twoproteins sharing structural similarity but differing in localization.This finding supports the emerging concept of synaptic vesicles as thesimplified and specialized form of a generic'trafflcking organelle.[Janz et al., J. Biol. Chem. 273(5):2851-2857 (1998)]. The sequence forcellugyrin derived from the Norway rat, Rattus norvegicus has beendeposited in the Genbank database on Dec. 23, 1997, designated accessionnumber AF039085. See also, Janz et al., J. Biol. Chem. 273 (1998), inpress.

[0087] Given the medical importance of synaptic transmission, effortsare currently being under taken to identify new, native proteins thatmay be part of a simplified and specialized generic traffickingorganelle in the form of synaptic vesicles. We describe herein theidentification of a novel polypeptide which has homology tosynaptogyrin, designated herein as PRO615.

[0088] 26. PRO618

[0089] Enteropeptidase is a key enzyme in the intestinal digestioncascade specifically cleaves the acidic propeptide from trypsinogen toyield active trypsin. This cleavage initiates a cascade of proteolyticreactions leading to the activation of many pancreatic zymogens.

[0090] See, for example, Matsushima et al., J. Biol. Chem.269(31):19976-19982 (1994), Kitamoto et al., Proc. Nat. Acad. Sci.,91(16):7588-7592 (1994). Enterokinase (enteropeptidase) is a related tomammalian serine proteases involved in digestion, coagulation, andfibrinolysis. LaVallie et al., J Biol Chem., 268(31):23311-23317 (1993).

[0091] Given the medical importance of digestive processes, efforts arecurrently being under taken to identify new, native proteins that may beinvolved in digestion, coagulation, and fibrinolysis. We describe hereinthe identification of a novel polypeptide which has homology toenteropeptidase, designated herein as PRO618.

[0092]27. PRO719

[0093] Lipoprotein lipase is a key enzyme that mediates the hydrolysisof triglycerides and phospholipids present in circulating plasmalipoproteins (Dugi et al., J. Biol. Chem. 270:25396-25401 (1995)).Moreover, lipoprotein lipase has been shown to mediate the uptake oflipoproteins into cells, wherein cellular uptake of lipoproteins isinitiated by binding of lipoprotein lipase to cell surface proteoglycansand to the low density lipoprotein (LDL) receptor-related protein (Krappet al., J. Lipid Res. 36:2362-2373 (1995)). Thus, it is clear thatlipoprotein lipase plays an extremely important role in lipoprotein andcholesterol metabolism. There is, therefore, substantial interest inidentifying novel polypeptides that share sequence homology and/orbiological activity with lipoprotein lipase. We herein describe theidentification and characterization of a novel polypeptide havingsequence homology to lipoprotein lipase H, designated herein as PRO719.

[0094] 28. PRO724

[0095] The low density lipoprotein (LDL) receptor is a membrane-boundprotein that plays a key role in cholesterol homeostasis, mediatingcellular uptake of lipoprotein particles by high affinity binding to itsligands, apolipoprotein (apo) B-100 and apoE. The ligand-binding domainof the LDL receptor contains 7 cysteine-rich repeats of approximately 40amino acids, wherein each repeat contains 6 cysteines, which form 3intra-repeat disulfide bonds. These unique structural features providethe LDL receptor with its ability to specifically interact with apoB-100 and apoE, thereby allowing for transport of these lipoproteinparticles across cellular membranes and metabolism of their components.Soluble fragments containing the extracellular domain of the LDLreceptor have been shown to retain the ability to interact with itsspecific lipoprotein ligands (Simmons et al., J. Biol. Chem.272:25531-25536 (1997)). Thus, it is clear that the LDL receptor isintimately involved in important physiological activities related tocholesterol metabolism. As such, there is substantial interest inidentifying novel LDL receptor homolog proteins. We herein describe theidentification and characterization of a novel polypeptide havinghomology to the human LDL receptor protein, designated herein as PRO724.

[0096] 29. PRO772

[0097] Expression of the human gene A4 is enriched in the colonicepithelium and is transcriptionally activated on differentiation ofcolonic epithelial cells in vitro (Oliva et al., Arch. Biochem. Biophys.302:183-192 (1993) and Oliva et al., Am. J. Physiol. 272: C957-C965(1997)). A4 cDNA contains an open reading frame that predicts apolypeptide of approximately 17 kilodaltons in size. Hydropathy analysisof the A4 protein revealed four putative membrane-spanningalpha-helices. Immunocytochemical studies of cells expressing A4 proteinindicated that expression is localized to the endoplasmic reticulum. Thefour membrane-spanning domains and the biophysical characteristics ofthe A4 protein suggest that it belongs to a family of integral membraneproteins called proteolipids, some of which multimerize to form ionchannels. In fact, preliminary evidence has suggested that A4 may itselfmultimerize and take on the properties of an ion channel (Oliva et al.,Am. J. Physiol. 272:C957-C965 (1997)). Given the importance of ionchannels in maintaining cellular homeostasis, there is a significantinterest in identifying novel polypeptides having homology to known andputative ion channels We herein describe the identification andcharacterization of a novel polypeptide having homology to the putativeion channel protein, A4, designated herein as PRO772.

[0098] 30. PRO852

[0099] Proteases are enzymatic proteins which are involved in a largenumber of very important biological processes in mammalian andnon-mammalian organisms. Numerous different protease enzymes from avariety of different mammalian and non-mammalian organisms have beenboth identified and characterized. The mammalian protease enzymes playimportant roles in many different biological processes including, forexample, protein digestion, activation, inactivation, or modulation ofpeptide hormone activity, and alteration of the physical properties ofproteins and enzymes.

[0100] In light of the important physiological roles played by proteaseenzymes, efforts are currently being undertaken by both industry andacademia to identify new, native protease homologs. Many of theseefforts are focused on the screening of mammalian recombinant DNAlibraries to identify the coding sequences for novel secreted andmembrane-bound receptor proteins. Examples of screening methods andtechniques are described in the literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)]. We herein describe the identification of novel polypeptideshaving homology to various protease enzymes, designated herein as PRO852polypeptides.

[0101] 31. PRO853

[0102] Studies have reported that the redox state of the cell is animportant determinant of the fate of the cell. Furthermore, reactiveoxygen species have been reported to be cytotoxic, causing inflammatorydisease, including tissue necrosis, organ failure, atherosclerosis,infertility, birth defects, premature aging, mutations and malignancy.Thus, the control of oxidation and reduction is important for a numberof reasons, including the control and prevention of strokes, heartattacks, oxidative stress, hypertension and may be associated with thedevelopment of malignancies. The levels of antioxidant enzymes, such asreductases, which catalyze the conversion of reactive oxygen species towater have been shown to be low in cancer cells. In particular,malignant prostate epithelium may have lowered expression of suchantioxidant enzymes [Baker et., Prostate 32(4):229-233 (1997)]. In thisregard, reductases, are of interest. In addition, the transcriptionfactors, NF-kappa B and AP-1, are known to be regulated by redox stateand to affect the expression of a large variety of genes thought to beinvolved in the pathogenesis of AIDS, cancer, atherosclerosis anddiabetic complications. Publications further describing this subjectmatter include Engman et al., Anticancer Res. (Greece), 17:4599-4605(1997), Kelsey, et al., Br. J. Cancer, 76(7):852-4 (1997); Friedrich andWeiss, J. Theor. Biol., 187(4):52940 (1997) and Pieulle, et al., J.Bacteriol., 179(18):5684-92 (1997). Given the physiological importanceof redox reactions in vivo, efforts are currently being under taken toidentify new, native proteins which are involved in redox reactions. Wedescribe herein the identification of a novel prostate specificpolypeptide which has sequence similarity to reductase, designatedherein as PRO853.

[0103] 32. PRO860

[0104] Neurofascin is a member of the LI subgroup of the cellularadhesion molecule (“CAM”) family of nervous system adhesion moleculesand is involved in cellular aggregation. Cell-cell recognition andpatterning of cell contacts have a critical role in mediating reversibleassembly of a wide variety or transcellular complexes in the nervoussystem. Cell interactions may be regulated through modulation of ankyrinbinding to neurofascin. See, for example, Tuvia et al., Proc. Nat Acad.Sci., 94(24) 12957-12962 (1997). Neurofascin has been described as amember of the L1 subgroup of the immunoglobulin superfamily implicatedin neurite extension during embryonic development for which numerousisoforms have been detected at various stages of development. See alsoHassel et al., J. Biol. Chem., 272(45) 28742-28749 (1997), Grumet.,Cell. Tissue Res. 290(2) 423-428 (1997), Garver et al., J. Cell. Biol.,137:703-714 (1997), and Lambert et al., J. Neurosci., 17:7025-7-36(1997).

[0105] Given the physiological importance of cellular adhesion moleculesand development of the nervous system in vivo, efforts are currentlybeing under taken to identify new, native proteins which are involved inregulation of cellular interactions in the nervous system. We describeherein the identification and characterization of a novel polypeptidewhich has sequence similarity to neurofascin, designated herein asPRO860.

[0106] 33. PRO846

[0107] The CMRF35 monoclonal antibody was used to identify a cellmembrane antigen, designated CMRF35, which is present on the surface ofmonocytes, neutrophils, a proportion of peripheral blood T and Blymphocytes and lymphocytic cell lines. The CMRF35 cDNA encodes a novelintegral membrane glycoprotein member of the immunoglobulin (Ig) genesuperfamily. The molecule comprises (a) a single extracellular Igvariable domain remarkably similar to the Fc receptor for polymeric IgAand IgM, (b) a membrane-proximal domain containing a high proportion ofproline, serine and threonine residues that was predicted to be heavilyO-glycosylated, (c) an unusual transmembrane anchor that contained aglutamic acid and a proline residue and (d) a short cytoplasmic tail.Transcripts encoding the CMRF35 protein have been detected in earlymonocytic cell lines, in peripheral blood T cells and in some Blymphoblastoid cell lines, confirming the results of immunocytologicalstaining. Jackson et al., Eur. J. Immunol. 22(5):1157-1163 (1992).CMRF-35 molecules are differentially expressed in hematopoietic cells,and the expression of the antigen was shown to be markedly influenced bystimulation with mitogens and cytokines. See, for example, Clark et al.,Exp. Hematol. 25(8):759 (1997), Daish et al., Immunol. 79(1):55-63(1993), and Clark et al., Tissue Antigens 48:461 (1996).

[0108] Given the physiological importance of the immune system andantigens associated with various immune system cells, efforts arecurrently being under taken to identify new, native proteins which areexpressed on various cells of the immune system. We describe herein theidentification of a novel polypeptide which has sequence similarity toCMRF35, designated herein as PRO846.

[0109] 34. PRO862

[0110] Lysozyme is a protein which is widely distributed in severalhuman tissues and secretions including milk, tears and saliva. It hasbeen demonstrated to hydrolyze linkages between N-acetylglucosamines. Ithas been demonstrated to be an inhibitor of chemotaxis and of theproduction of toxic oxygen free radicals and may also have some role inthe calcification process. As such, there is substantial interest inidentifying novel polypeptides having homology to lysozyme. We describeherein the identification of a novel polypeptide which has sequencesimilarity to lysozyme.

[0111] 35. PRO864

[0112] Wnt-4 is a secreted glycoprotein which correlates with, and isrequired for, kidney tubulogenesis. Mice lacking Wnt-4 activity fail toform pretubular cell aggregates; however, other aspects of mesenchymaland ureteric development are unaffected. Thus, Wnt-4 appears to act asan autoinducer of the mesenchyme to epithelial transition that underliesnephron development. Stark et al., Nature ;372(6507):679-683 (1994). Inaddition, members of the Wnt gene family code for cysteine-rich,secreted proteins, which are differentially expressed in the developingbrain and possibly act as intercellular signaling molecules. A Wnt gene,e.g., Wnt-1 is known to be essential for specification of the midbraincell fate. Yoshioka et al., Biochem. Biophys. Res. Commun.203(3):1581-1588 (1994). Several member of the Wnt family of secretedfactors are strongly implicated as regulators of mammary cellular growthand differentiation. Shimizu et al., Cell Growth Differ. 8(12)1349-1358. Wnt-4 is normally expressed in early pregnancy. Wnt-4 maytherefore be a local signal driving epithelial branching in pregnancy.Edwards P A, Biochem Soc Symp.63:21-34 (1998). See also, Lipschutz J H,Am. J. Kidney Dis. 31(3):383-397, (1998). We describe herein theidentification and characterizaton of a novel polypeptide which hassequence similarity to Wnt-4, designated herein as PRO864.

[0113] 36. PRO792

[0114] At least two cell-derived signals have been shown to be necessaryfor the induction of immunoglobulin isotype switching in Bells. Thefirst signal is given by either of the soluble lymphokines, interleukin(IL)4 or IL-I 3, which induce germline epsilon transcript expression,but this alone is insufficient to trigger secretion of immunoglobulin E(IgE). The second signal is provided by a physical interaction betweenB-cells and activated Tells, basophils and mast cells, and it has beenshown that the CD40ICD40 ligand pairing is crucial for mediating IgEsynthesis. Additionally, amongst the numerous pairs of surface adhesionmolecules that are involved in IgE synthesis, the CD23/CD21 pair appearsto play a key role in the generation of IgE. CD23 is a protein that ispositively and negatively regulated by factors which increase ordecrease IgE production, respectively. Antibodies to CD23 have beenshown to inhibit IL-4-induced human IgE production in vitro and toinhibit antigen-specific IgE responses in a rat model, in an isotypeselective manner (Bonnefoy et al., Eur. Respir. J. Suppl. 22:63S66S(1996)). CD23 interacts with CD21 on B-cells, preferentially driving IgEproduction. Given that the CD23 protein plays an extremely importn rolein the induction of a mammalian IgE response, there is significantinterest in identifying novel polypeptides having homology to CD23. Weherein describe the identification and characterization of a novelpolypeptide having homology to CD23, designated herein as PRO792.

[0115] 37. PRO866

[0116] Mindin and spondin proteins are secreted proteins that arestructurally related to one another and which have been identified in avariety of organisms. For example, Higashijima et al., Dev Biol.192:211-227 (1997) have reported the identification of spondin andmindin expression in floor plate cells in the zebrafish embryonic axis,thereby suggesting that mindin and spondin proteins play important rolesin embryonic development. This same group has reported that mindin andspondin proteins function as extracellular matrix proteins that have ahigh affinity for the basal lamina. (Id.). It has been reported thatF-spondin is a secreted protein that promotes neural adhesion andneurite extension (Klar et al., Cell 69:95-110 (1992) and that M-spondinis an extracellular matrix protein that localizes to muscle attachmentsites in Drosophila (Umemiya et al., Dev. Biol. 186:165-176 (1997)).Thus, there is significant interest in identifying novel polypeptideshaving homology to the mindin and spondin proteins. We herein describethe identification and characterization of a novel polypeptide havinghomology to mindin2 and mindin1, designated herein as PRO866.

[0117] 38. PRO871

[0118] Cyclophilins are a family of proteins that bind to cyclosporin Aand possess peptidyl-prolyl cis-trans isomerase activity (Sherry et al.,Proc. Natl. Acad. Sci. USA 95:1758-1763 (1998)). In addition,cyclophilins are secreted by activated cells and act in a cytokine-likemanner, presumably via signaling through a cell surface cyclophilinreceptor. Host cell-derived cyclophilin A has been shown to beincorporated into HIV-1 virions and its incorporation has been shown tobe essential for viral infectivity. Thus, one or more the cyclophilinsmay be directly associated with HIV-1 infectivity. Given the obviousimportance of the cyclophilin proteins, there is substantial interest inidentifying novel polypeptides which have sequence homology to one ormore of the cyclophilin proteins. We herein describe the identificationand characterization of a novel polypeptide having homology tocyclophilin-like protein CyP-60, designated herein as PRO871.

[0119] 39. PRO873

[0120] Enzymatic proteins play important roles in the chemical reactionsinvolved in the digestion of foods, the biosynthesis of macromolecules,the controlled release and utilization of chemical energy, and otherprocesses necessary to sustain life. Enzymes have also been shown toplay important roles in combating various diseases and disorders. Forexample, liver carboxylesterases have been reported to assist insensitizing human tumor cells to the cancer prodrugs. Danks et al.,report that stable expression of the cDNA encoding a carboxylesterase inRh30 human rhabdomyosarcoma cells increased the sensitivity of the cellsto the CPT-11 cancer prodrug 8.1-fold. Cancer Res. (1998) 58(1):20-22.The authors propose that this prodrug/enzyme combination could beexploited therapeutically in a manner analogous to approaches currentlyunder investigation with the combinations of ganciclovir/herpes simplexvirus thymidine kinase and 5-fluorocytosine/cytosine deaminase. van Peltet al. demonstrated that a 55 kD human liver carboxylesterase inhibitsthe invasion of Plasmodium falciparum malaria sporozoites into primaryhuman hepatocytes in culture. J Hepatol (1997) 27(4):688-698.

[0121] Carboxylesterases have also been found to be of importance in thedetoxification of drugs, pesticides and other xenobiotics. Purifiedhuman liver carboxylesterases have been shown to be involved in themetabolism of various drugs including cocaine and heroin. Prindel et al.describe the purification and cloning of a broad substrate specificityhuman liver carboxylesterase which catalyzes the hydrolysis of cocaineand heroin and which may play an important role in the degradation ofthese drugs in human tissues. J. Biol. Chem. (1997)6:272(23):14769-14775. Brzenzinski et al. describe a spectrophotometriccompetitive inhibition assay used to identify drug or environmentalesters that are metabolized by carboxylesterases. Drug Metab Dispos(1997) 25(9):1089-1096.

[0122] In light of the important physiological roles played bycarboxylesterases, efforts are being undertaken by both industry andacademia to identify new, native carboxylesterase homologs. We hereindescribe the identification and characterization of a novel polypeptidehaving homology to carboxylesterase, designated herein as PRO873.

[0123] 40. PRO940

[0124] CD33 is a cell-surface protein that is a member of thesialoadhesin family of proteins that are capable of mediatingsialic-acid dependent binding with distinct specificities for both thetype of sialic acid and its linkage to subterminal sugars. CD33 isspecifically expressed in early myeloid and some monocyte cell lineagesand has been shown to be strongly associated with various myeloid tumorsincluding, for example, acute non-lymphocytic leukemia (ANLL). As such,CD33 has been suggested as a potential target for the treatment ofcancers associated with high level expression of the protein. There is,therefore, significant interest in the identification of novelpolypeptides having homology to CD33. In fact, one CD33 homolog(designated CD33L) has already been identified and described (see Takeiet al., Cytogenet. Cell Genet. 78:295-300 (1997)). We herein describethe identification of another novel polypeptide having homology to CD33,designated herein as PRO940. The novel polypeptide described herein alsoexhibits significant homology to the human OB binding proteinsdesignated HSU71382_(—)1 and HSU71383_(—)1 in the Dayhoff database(version 35.45 SwissProt 35).

[0125] 41. PRO941

[0126] Cadherins are a large family of transmembrane proteins. Cadherinscomprise a family of calcium-dependent glycoproteins that function inmediating cell-cell adhesion in virtually all solid tissues ofmulticellular organisms. At least cadherins 1-13 as well as types B, T,EP, M, N, P and R have been identified and characterized. Among thefunctions cadherins are known for, with some exceptions, are thatcadherins participate in cell aggregation and are associated withcell-cell adhesion sites. Recently, it has been reported that while allcadherins share multiple repeats of a cadherin specific motif believedto correspond to folding of extracellular domains, members of thecadherin superfamily have divergent structures and, possibly, functions.In particular it has been reported that members of the cadherinsuperfamily are involved in signal transduction. See, Suzuki, J. CellBiochem., 61(4):531-542 (1996). Cadherins are further described inTanihara et al., J. Cell Sci., 107(6):1697-1704 (1994), Aberle et al.,J. Cell Biochem., 61(4):514-523 (1996) and Tanihara et al., Cell Adhes.Commun., 2(1):15-26 (1994). We herein describe the identification andcharacterization of a novel polypeptide having homology to a cadherinprotein, designated herein as PRO941.

[0127] 42. PRO944

[0128] Clostridium perfringens enterotoxin (CPE) is considered to be thevirulence factor responsible for causing the symptoms of C. perfringenstype A food poisoning and may also be involved in other human andveterinary illnesses (McClane, Toxicon. 34:1335-1343 (1996)). CPEcarries out its adverse cellular functions by binding to anapproximately 50 kD cell surface receptor protein designated theClostridium perfringens enterotoxin receptor (CPE-R) to form anapproximately 90,000 kD complex on the surface of the cell. cDNAsencoding the CPE-R protein have been identified characterized in bothhuman and mouse (Katabira et al., J. Cell Biol. 136:1239-1247 (1997) andKatabira et al., J. Biol. Chem. 272:26652-26658 (1997)). Since the CPEtoxin has been reported to cause a variety of illnesses in mammalianhosts and those illnesses are initiated by binding of the CPE toxin tothe CPE-R, there is significant interest in identifying novel CPE-Rhomologs. We herein describe the identification and characterization ofa novel polypeptide having homology to the CPE-R, designated herein asPRO944.

[0129] 43. PRO983

[0130] Membrane-bound proteins include not only cell-surfacemembrane-bound proteins, but also proteins that are found on the surfaceof intracellular vesicles. These vesicles are involved in exocytosis,which is the fusion of secretory vesicles with the cellular plasmamembrane, and have two main functions. One is the discharge of thevesicle contents into the extracellular space, and the second is theincorporation of new proteins and lipids into the plasma membraneitself. Exocytosis can be either constitutive or regulated. Alleukaryotic cells exhibit constitutive exocytosis, which is marked by theimmediate fusion of the secretory vesicle after formation. In contrast,regulated exocytosis results in the accumulation of the secretoryvesicles that fuse with the plasma membrane upon receipt of anappropriate signal by vesicle-associated membrane proteins. Usually,this signal is an increase in the cytosolic free Ca²⁺ concentration.However, regulated exocytosis that is independent of Ca²⁺ has beenreported (see, e.g. Fujita-Yoshigaki et al. J. Biol. Chem. (1996)31:271(22):13130-13134). Regulated exocytosis is chemical to manyspecialized cells, including neurons (neurotransmitter release fromsynaptic vesicles), adrenal chromaffin cells (adrenaline secretion),pancreatic acinar cells (digestive enzyme secretion), pancreatic β-cells(insulin secretion), mast cells (histamine secretion), mammary cells(milk protein secretion), sperm (enzyme secretion), egg cells (creationof fertilization envelope) and adipocytes (insertion of glucosetransporters into the plasma membrane).

[0131] Disorders involving exocytosis are known. For example,inflammatory mediator release from mast cells leads to a variety ofdisorders, including asthma. Similarly, Chediak-Higashi Syndrome (CHS)is a rare autosomal recessive disease in which neutrophils, monocytesand lymphocytes contain giant cytoplasmic granules. Accordingly, theproteins involved in exocytosis are of paramount interest and effortsare being undertaken by both industry and academia to identify new,vesicle-associated proteins. For example, Skehel et al. identified a33-kilodalton membrane protein in Aplysia, termed VAP-33, which isrequired for the exocytosis of neurotransmitter. Science (1995)15:269(5230):1580-1583, and Neuropharmacology (1995) 34(11):1379-1385.Many efforts are focused on the screening of mammalian recombinant DNAlibraries to identify the coding sequences for novel vesicle- associatedmembrane proteins. It is an object of the invention to provide proteinshaving homology to the vesicle associated protein, VAP-33, designatedherein as PRO983.

[0132] 44. PRO1057

[0133] Proteases are enzymatic proteins which are involved in a largenumber of very important biological processes in mammalian andnon-mammalian organisms. Numerous different protease enzymes from avariety of different mammalian and non-mammalian organisms have beenboth identified and characterized. The mammalian protease enzymes playimportant roles in many different biological processes including, forexample, protein digestion, activation, inactivation, or modulation ofpeptide hormone activity, and alteration of the physical properties ofproteins and enzymes.

[0134] In light of the important physiological roles played by proteaseenzymes, efforts are currently being undertaken by both industry andacademia to identify new, native protease homologs. Many of theseefforts are focused on the screening of mammalian recombinant DNAlibraries to identify the coding sequences for novel secreted proteins.Examples of screening methods and techniques are described in theliterature [see, for example, Klein et al., Proc. Natl. Acad. Sci.,93:7108-7113 (1996); U.S. Pat. No. 5,536,637)]. We herein describe theidentification of novel polypeptides having homology to various proteaseenzymes, designated herein as PRO1057 polypeptides.

[0135] 45. PRO1071

[0136] Thrombospondin-1 is a trimeric high molecular weight glycoproteinthat is released from platelet alpha-granules in response to thrombinstimulation and that is also a transient component of the extracellularmatrix in developing and repairing tissues (Adams, Int. J. Biochem. CellBiol. 29:861-865 (1997) and Qian et al., Proc. Soc. Exp. Biol. Med.212:199-207 (1996)). A variety of factors regulate thrombospondinexpression and the protein is degraded by both extracellular andintracellular routes. Thrombospondin-I functions as a cell adhesionmolecule and also modulates cell movement, cell proliferation, neuriteoutgrowth and angiogenesis. As such, there is substantial interest inidentifying novel polypeptides having homology to thrombospondin. Weherein describe the identification and characterization of a novelpolypeptide having homology to thrombospondin, designated herein asPRO1071.

[0137] 46. PRO1072

[0138] Studies have reported that the redox state of the cell is animportant determinant of the fate of the cell. Furthermore, reactiveoxygen species have been reported to be cytotoxic, causing inflammatorydisease, including tissue necrosis, organ failure, atherosclerosis,infertility, birth defects, premature aging, mutations and malignancy.Thus, the control of oxidation and reduction is important for a numberof reasons, including the control and prevention of strokes, heartattacks, oxidative stress, hypertension and may be associated with thedevelopment of malignancies. The levels of antioxidant enzymes, such asreductases, which catalyze the conversion of reactive oxygen species towater have been shown to be low in cancer cells. In particular,malignant prostate epithelium may have lowered expression of suchantioxidant enzymes [Baker et al., Prostate 32(4):229-233 (1997)]. Inthis regard, reductases, are of interest. In addition, the transcriptionfactors, NF-kappa B and AP-1, are known to be regulated by redox stateand to affect the expression of a large variety of genes thought to beinvolved in the pathogenesis of AIDS, cancer, atherosclerosis anddiabetic complications. Publications further describing this subjectmatter include Engman et al., Anticancer Res. (Greece), 17:4599-4605(1997), Kelsey, et al., Br. J. Cancer, 76(7):852-854 (1997); Friedrichand Weiss, J. Theor. Biol., 187(4):52940 (1997) and Pieulle, et al., J.Bacteriol., 179(18):5684-92 (1997). Given the physiological importanceof redox reactions in vivo, efforts are currently being under taken toidentify new, native proteins which are involved in redox reactions. Wedescribe herein the identification of a novel polypeptide which hassequence similarity to reductase enzymes, designated herein as PRO1072.

[0139] 47. PRO1075

[0140] Protein disulfide isomerase is an enzymatic protein which isinvolved in the promotion of correct refolding of proteins through theestablishment of correct disulfide bond formation. Protein disulfideisomerase was initially identified based upon its ability to catalyzethe renaturation of reduced denatured RNAse (Goldberger et al., J. Biol.Chem. 239:1406-1410 (1964) and Epstein et al., Cold Spring Harbor Symp.Quant. Biol. 28:439-449 (1963)). Protein disulfide isomerase has beenshown to be a resident enzyme of the endoplasmic reticulum which isretained in the endoplasmic reticulum via a −KDEL or −HDEL amino acidsequence at its C-terminus.

[0141] Given the importance of disulfide bond-forming enzymes and theirpotential uses in a number of different applications, for example inincreasing the yield of correct refolding of recombinantly producedproteins, efforts are currently being undertaken by both industry andacademia to identify new, native proteins having homology to proteindisulfide isomerase. Many of these efforts are focused on the screeningof ii mammalian recombinant DNA libraries to identify the codingsequences for novel protein disulfide isomerase homologs. Examples ofscreening methods and techniques are described in the literature [see,for example, Klein et al., Proc. Natl. Acad. Sci., 93:7108-7113 (1996);U.S. Pat. No. 5,536,637)]. We herein describe a novel polypeptide havinghomology to protein disulfide isomerase, designated herein as PRO1075.

[0142] 48. PRO181

[0143] In Drosophila, the dorsal-ventral polarity of the egg chamberdepends on the localization of the oocyte nucleus and the gurken RNA tothe dorsal-anterior corner of the oocyte. Gurken protein presumably actsas a ligand for the drosophila EGF receptor (torpedo/DER) expressed inthe somatic follicle cells surrounding the oocyte. Cornichon is a generequired in the germline for dorsal-ventral signaling (Roth et al., Cell81:967-978 (1995)). Cornichon, gurken and torpedo also function in anearlier signaling event that establishes posterior follicle cell fatesand specifies the anterior-posterior polarity of the egg chamber.Mutations in any or all of these genes prevent the formation of acorrectly polarized microtubule cytoskeleton required for properlocalization of the anterior and posterior determinants bicoid and oskarand for the asymmetric positioning of the oocyte nucleus. Thus, it isclear that the cornichon gene product plays an important role in earlydevelopment. We herein describe the identification and characterizationof a novel polypeptide having homology to the cornichon protein,designated herein as PRO181.

[0144] 49. PRO195

[0145] Efforts are currently being undertaken to identify andcharacterize novel transmembrane proteins. We herein describe theidentification and characterization of a novel transmembranepolypeptide, designated herein as PRO195.

[0146] 50. PRO865

[0147] Efforts are currently being undertaken to identify andcharacterize novel secreted proteins. We herein describe theidentification and characterization of a novel secreted polypeptide,designated herein as PRO865.

[0148] 51. PRO827

[0149] VLA-2 is an cell-surface integrin protein that has beenidentified and characterized in a number of mammalian organisms,including both mouse and human. VLA-2 has been shown to be a receptor onthe surface of cells for echovirus-1 (EV-1) which mediates infection ofVLA-2-expressing cells by EV-1 (Zhang et al., Virology 235(2):293-301(1997) and Bergelson et al., Science 255:1718-1720 (1992)). VLA-2 hasalso been shown to mediate the interaction of collagen with endotheliumduring in vitro vascular tube formation (Jackson et al., Cell Biol. Int.18(9):859-867 (1994)). Various other integrin proteins that sharevarious degrees of amino acid sequence homology with VLA-2 have beenidentified and characterized in a variety of mammalian organism. Theseintegrins have been reported to play important roles in a variety ofdifferent physiological functions. Therefore, there is significantinterest in identifying novel polypeptides having homology to one ormore of the integrin proteins. We herein describe the identification andcharacterization of a novel polypeptide having homology to VLA-2integrin protein, designated herein as PRO827.

[0150] 52. PRO1114

[0151] Many important cytokine proteins have been identified andcharacterized and shown to signal through specific cell surface receptorcomplexes. For example, the class II cytokine receptor family (CRF2)includes the interferon receptors, the interleukin-10 receptor and thetissue factor CRFB4 (Spencer et al., J. Exp. Med. 187:571-578 (1998) andKotenko et al., EMBO J. 16:5894-5903 (1997)). Thus, the multitude ofbiological activities exhibited by the various cytokine proteins isabsolutely dependent upon the presence of cytokine receptor proteins onthe surface of target cells. There is, therefore, a significant interestin identifying and characterizing novel polypeptides having homology toone or more of the cytokine receptor family. We herein describe theidentification and characterization of a novel polypeptide havinghomology to cytokine receptor family-4 proteins, designated herein asPRO1117.

[0152] Interferons (IFNs) encompass a large family of secreted proteinsoccurring in vertebrates. Although they were originally named for theirantiviral activity, growing evidence supports a critical role for IFNsin cell growth and differentiation (Jaramillo et al., CancerInvestigation 13(3):327-338 (1995)). IFNs belong to a class of negativegrowth factors having the ability to inhibit the growth of a widevariety of cells with both normal and transformed phenotypes. IFNtherapy has been shown to be beneficial in the treatment of humanmalignancies such as Karposi's sarcoma, chronic myelogenous leukemia,non-Hodgkin's lymphoma, and hairy cell leukemia as well as in thetreatment of infectious diseases such as hepatitis B (Gamliel et al.,Scanning Microscopy 2(1):485-492 (1988), Einhorn et al., Med. Oncol. &Tumor Pharmacother. 10:25-29 (1993), Ringenberg et al., MissouriMedicine 85(1):21-26 (1988), Saracco et al., Journal of Gastroenterologyand Hevatology 10:668-673 (1995), Gonzalez-Mateos et al.,Hepato-Gastroenterology 42:893-899 (1995) and Malaguarnera et al.,Pharmacotherapy 17(5):998-1005 (1997)).

[0153] Interferons can be classified into two major groups based upontheir primary sequence. Type I interferons, IFN-α and IFN-β, are encodedby a superfamily of intronless genes consisting of the IFN-α gene familyand a single IFN-β gene that are thought to have arisen from a commonancestral gene. Type I interferons may be produced by most cell types.Type II IFN, or IFN-γ, is restricted to lymphocytes (F cells and naturalkiller cells) and is stimulated by nonspecific T cell activators orspecific antigens in vivo.

[0154] Although both type I and type II IFNs produce similar antiviraland antiproliferative effects, they act on distinct cell surfacereceptors, wherein the binding is generally species specific (Langer etal., Immunol. Today 9:393-400 (1988)). Both IFN-α and IFN-β bindcompetitively to the same high affinity type I receptor, whereas IFN-γbinds to a distinct type II receptor. The presence and number of IFNreceptors on the surface of a cell does not generally reflect thesensitivity of the cell to IFN, although it is clear that the effects ofthe IFN protein is mediated through binding to a cell surface interferonreceptor. As such, the identification and characterization of novelinterferon receptor proteins is of extreme interest.

[0155] We herein describe the identification and characterization ofnovel interferon receptor polypeptides, designated herein as “PRO1114interferon receptor” polypeptides. Thus, the PRO1114 polypeptides of thepresent invention represents a novel cell surface interferon receptor.

[0156] 53. PRO237

[0157] Carbonic anhydrase is an enzymatic protein that which aids carbondioxide transport and release in the mammalian blood system bycatalyzing the synthesis (and the dehydration) of carbonic acid from(and to) carbon dioxide and water. Thus, the actions of carbonicanhydrase are essential for a variety of important physiologicalreactions in the mammal. As such, there is significant interest in theidentification and characterization of novel polypeptides havinghomology to carbonic anhydrase. We herein describe the identificationand characterization of a novel polypeptide having homology to carbonicanhydrase, designated herein as PRO237.

[0158] 54. PRO541

[0159] Numerous trypsin inhibitory proteins have been identified andcharacterized (see, e.g., Yamakawa et al., Biochim. Biophys. Acta1395:202-208 (1998) and Mizukl et al., Mammalian Genome 3:274-280(1992)). Trypsin inhibitor proteins play important roles in a variety ofdifferent physiological and biological pathways and are specificallyinvolved in such processes as the regulation of protein degradation,digestion, and the like. Given the important roles played by suchenzymatic proteins, there is significant interest in identifying andcharacterizing novel polypeptides having homology to known trypsininhibitor proteins. We herein describe the identification andcharacterization of a novel polypeptide having homology to a trypsininhibitor protein, designated herein as PRO541.

[0160] 55. PRO273

[0161] Leukocytes include monocytes, macrophages, basophils, andeosinophils and play an important role in the immune response. Thesecells are important in the mechanisms initiated by T and/or Blymphocytes and secrete a range of cytokines which recruit and activateother inflammatory cells and contribute to tissue destruction.

[0162] Thus, investigation of the regulatory processes by whichleukocytes move to their appropriate destination and interact with othercells is critical. Currently, leukocytes are thought to move from theblood to injured or inflamed tissues by rolling along the endothelialcells of the blood vessel wall. This movement is mediated by transientinteractions between selectins and their ligands. Next, the leukocytemust move through the vessel wall and into the tissues. This diapedesisand extravasation step involves cell activation which promotes a morestable leukocyte-endothelial cell interaction, again mediated byintegrins and their ligands.

[0163] Chemokines are a large family of structurally related polypeptidecytokines. These molecules stimulate leukocyte movement and may explainleukocyte trafficking in different inflammatory situations. Chemokinesmediate the expression of particular adhesion molecules on endothelialcells, and they produce chemoattractants which activate specific celltypes. In addition, the chemokines stimulate proliferation and regulateactivation of specific cell types. In both of these activities,chemokines demonstrate a high degree of target cell specificity.

[0164] The chemokine family is divided into two subfamilies based onwhether two amino terminal cysteine residues are immediately adjacent(C-C) or separated by one amino acid (C-X-C). Chemokines of the C-X-Cfamily generally activate neutrophils and fibroblasts while the C-Cchemolines act on a more diverse group of target cells includingmonocytes/macrophages, basophils, eosinophils and T lymphocytes. Theknown chemokines of both subfamilies are synthesized by many diversecell types as reviewed in Thomson A. (1994) The Cytokine Handbook, 2 dEd. Academic Press, N.Y. Chemokines are also reviewed in Schalt T J(1994) Chemotactic Cytokines: Targets for Therapeutic Development.International Business Communications, Southborough Mass. pp 180-270;and in Paul W E (1993) Fundamental Immunology, 3rd Ed. Raven Press, N.Y.pp 822-826.

[0165] Known chemokines of the C-X-C subfamily include macrophageinflammatory proteins alpha and beta (MIP-1 and MIP-2), interleukin-8(IL-8), and growth regulated protein (GRO-alpha and beta).

[0166] MIP-2 was first identified as a 6 kDa heparin binding proteinsecreted by the mouse macrophage cell line RAW 264.7 upon stimulationwith lipopolysaccharide (LPS). MIP-2 is a member of the C-X-C (or CXC)subfamily of chemokines. Mouse MIP-2 is chemotactic for humanneutrophils and induces local neutrophil infiltration when injected intothe foot pads of mice. Rat MIP-2 shows 86% amino acid homology to themouse MIP-2 and is chemotactic for rat neutrophils but does notstimulate migration of rat alveolar macrophages or human peripheralblood eosinophils or lymphocytes. In addition, the rat M[P-2 has beenshown to stimulate proliferation of rat alveolar epithelial cells butnot fibroblasts.

[0167] Current techniques for diagnosis of abnormalities in inflamed ordiseased issues mainly rely on observation of clinical symptoms orserological analyses of body tissues or fluids for hormones,polypeptides or various metabolites. Problems exist with thesediagnostic techniques. First, patients may not manifest clinicalsymptoms at early stages of disease. Second, serological tests do notalways differentiate between invasive diseases and genetic syndromes.Thus, the identification of expressed chemokines is important to thedevelopment of new diagnostic techniques, effective therapies, and toaid in the understanding of molecular pathogenesis.

[0168] To date, chemokines have been implicated in at least thefollowing conditions: psoriasis, inflammatory bowel disease, renaldisease, arthritis, immune-mediated alopecia, stroke, encephalitis, MS,hepatitis, and others. In addition, non-ELR-containing chemokines havebeen implicated in the inhibition of angiogenesis, thus indicating thatthese chemokines have a rule in tumor vascularization and tumorigenesis.

[0169] Therefore it is the object of this invention to identifypolypeptides and nucleic acids encoding the same which have sequenceidentity and similarity with cytokine-induced neutrophilchemoattractants, MIP-1, MIP-2, and other related proteins. The effortsof this object are provided herein.

[0170] 56. PRO701

[0171] Beta neurexins and neuroligins are plasma membrane proteins thatare displayed on the neuronal cell surface. Neuroligin 1 is enriched insynaptic plasma membranes and acts as a splice site-specific ligand forbeta neurexins as described in Ichtchenko, et al., Cell, 81(3):435-443(1995). The extracellular sequence of neuroligin 1 is composed of acatalytically inactive esterase domain homologous toacetylcholinesterase. Neuroligin 2 and 3 are similar in structure andsequence to neuroligin 1. All neuroligins contain an N-terminalhydrophobic sequence with the characteristics of a cleaved signalpeptide followed by a large esterase homology domain, a highly conservedsingle transmembrane region, and a short cytoplasmic domain. The threeneuroligins are alternatively spliced at the same position and areexpressed at high levels only in the brain. Tight binding of the threeneuroligins to beta neurexins is observed only for beta neurexinslacking an insert in splice site 4. Thus, neuroligins constitute amultigene family of brain-specific proteins with distinct isoforms thatmay have overlapping functions in mediating recognition processesbetween neurons, see Ichtchenko, et al., J. Biol. Chem.,271(5):2676-2682 (1996). Moreover, neurexins and neuroligins have beenreported as functioning as adhesion molecules in a Ca²⁺ dependentreaction that is regulated by alternative splicing of beta neurexins,i.e., see Nguyen and Sudhof, J. Biol. Chem., 272(41):26032-26039 (1997).Given the foregoing, membrane bound proteins are of interest. Moregenerally, membrane-bound proteins and receptors can play an importantrole in the formation, differentiation and maintenance of multicellularorganisms. The fate of many individual cells, e.g., proliferation,migration, differentiation, or interaction with other cells, istypically governed by information received from other cells and/or theimmediate environment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor.

[0172] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0173] Efforts are being undertaken by both industry and academia toidentify new, native membrane-bound receptor proteins, particularlythose having sequence identity and/or similarity with neuroligins 1, 2and 3. Many efforts are focused on the screening of mammalianrecombinant DNA libraries to identify the coding sequences for novelsecreted and membrane-bound receptor proteins. Examples of screeningmethods and techniques are described in the literature [see, forexample, Klein et al., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S.Pat. No. 5,536,637)]. The results of such efforts are provided herein.

[0174] 57. PRO704

[0175] VIP36 is localized to the Golgi apparatus and the cell surface,and belongs to a family of legume lectin homologues in the animalsecretory pathway that might be involved in the trafficking ofglycoproteins, glycolipids, or both. It is further believed that VIP36binds to sugar residues of glycosphingolipids and/orgycosylphosphatidyl-inositol anchors and might provide a link betweenthe extracellular/luminal face of glycolipid rafts and the cytoplasmicprotein segregation machinery. Further regarding VIP36, it is believedthat there is a signal at its C-terminus that matches an internalizationconsensus sequence which confers its ability to cycle between the plasmamembrane and Golgi. See, Fiedler, et al, EMBO J., 13(7):1729-1740(1994); Fiedler and Simons, J. Cell Sci., 109(1):271-276 (1996); Itin,et al., MBO J., 14(10):2250-2256 (1995). It is believed that VIP36 iseither the same as or very closely related to the human GP36b protein.VIP36 and/or GP36b are of interest.

[0176] More generally, vesicular, cytoplasmic, extracellular andmembrane-bound proteins play important roles in the formation,differentiation and maintenance of multicellular organisms. The fate ofmany individual cells, e.g., proliferation, migration, differentiation,or interaction with other cells, is typically governed by informationreceived from other cells and/or the immediate environment. Thisinformation is often transmitted by secreted polypeptides (for instance,mitogenic factors, survival factors, cytotoxic factors, differentiationfactors, neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Thesesecreted polypeptides or signaling molecules normally pass through thecellular secretory pathway to reach their site of action in theextracellular environment, usually at a membrane-bound receptor protein.

[0177] Secreted proteins have various industrial applications, includinguse as pharmaceuticals, diagnostics, biosensors and bioreactors. Infact, most protein drugs available at present, such as thrombolyticagents, interferons, interleukins, erythropoietins, colony stimulatingfactors, and various other cytokines, are secretory proteins. Theirreceptors, which are membrane-bound proteins, also have potential astherapeutic or diagnostic agents. Receptor immunoadhesins, for instance,can be employed as therapeutic agents to block receptor-ligandinteraction. Membrane-bound proteins can also be employed for screeningof potential peptide or small molecule inhibitors of the relevantreceptor/ligand interaction. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. Transduction of signals that regulate cell growth anddifferentiation is regulated in part by phosphorylation of variouscellular proteins. Protein tyrosine kinases, enzymes that catalyze thatprocess, can also act as growth factor receptors. Examples includefibroblast growth factor receptor and nerve growth factor receptor.

[0178] Efforts are being undertaken by both industry and academia toidentify new, native vesicular, cytoplasmic, secreted and membrane-boundreceptor proteins, particularly those having sequence identity and/orsimilarity with VIP36. Many efforts are focused on the screening ofmammalian recombinant DNA libraries to identify the coding sequences fornovel secreted and membrane-bound receptor proteins. Examples ofscreening methods and techniques are described in the literature [see,for example, Klein et al., Proc. Natl. Acad. Sci., 93:7108-7113 (1996);U.S. Pat. No. 5,536,637)].

[0179] 58. PRO706

[0180] Acid phophatase proteins are secreted proteins whichdephophorylate terminal phosphate groups under acidic pH conditions.Acid phophatases contain a RHGXRXP amino acid sequence, which ispredicted to be mechanistically significant. Acid phosphatases may haveimportant functions in the diagnosis and treatment of human diseases.For example, prostatic acid phosphatase is a secreted protein uniquelyexpressed in prostatic tissue and prostate cancer. The level ofprostatic acid phosphatase is a potential prognostic factor for localand biochemical control in prostate cancer patients treated withradiotherapy, as described in Lankford et al., Int. J. Radiat. Oncol.Biol. Phys. 38(2):327-333 (1997). Research suggests that a cellularimmune response to prostatic acid phosphatase may mediate destructiveautoimmune prostatitis, and that xenogeneic forms of prostatic acidphosphatase may prove useful for immunotherapy of prostate cancer. SeeFong et al., J. Immunol. 169(7):3113-3117(1997). Seminal prostatic acidphosphatase levels correlate significantly with very low sperm levels(oligospermia) in individuals over 35, see Singh et al., Singapore Med.J. 37(6):598-599 (1996). Thus, prostatic acid phosphatase has beenimplicated in a variety of human diseases, and may have an importantfunction in diagnosis and therapy of these diseases. A series ofaminobenzylphosphatic acid compounds are highly potent inhibitors ofprostatic acid phosphatase, as described in Beers et al., Bioorg. Med.Chem. 4(10): 1693-1701 (1996).

[0181] More generally, extracellular proteins play an important role inthe formation, differentiation and maintenance of multicellularorganisms. The fate of many individual cells, e.g., proliferation,migration, differentiation, or interaction with other cells, istypically governed by information received from other cells and/or theimmediate environment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. These secreted polypeptides or signalingmolecules normally pass through the cellular secretory pathway to reachtheir site of action in the extracellular environment.

[0182] Secreted proteins have various industrial applications, includingpharmaceuticals, diagnostics, biosensors and bioreactors. Most proteindrugs available at present, such as thrombolytic agents, interferons,interleukins, erythropoietins, colony stimulating factors, and variousother cytokines, are secretory proteins. Their receptors, which aremembrane proteins, also have potential as therapeutic or diagnosticagents. Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins, particularly those havingsequence identity with prostate acid phosphatase precursor and lysosomalacid phosphatase precursor and in some cases, those having identity withDNA found in fetal heart. Many efforts are focused on the screening ofmammalian recombinant DNA libraries to identify the coding sequences fornovel secreted proteins. Examples of screening methods and techniquesare described in the literature [see, for example, Klein et al., Proc.Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No. 5,536,637)].

[0183] 59. PRO707

[0184] Cadherins are a large family of transmembrane proteins. At leastcadherins 1-13 as well as types B, E, EP, M, N, P and R have beencharacterized. Among the functions cadherins are known for, with someexceptions, cadherins participate in cell aggregation and are associatedwith cell-cell adhesion sites. Cadherins are further described inTanihara, et al., J. Cell Sci., 107(6):1697-1704 (1994) and Tanihara, etal., Cell Adhes. Commun., 2(1):15-26 (1994). Moreover, it has beenreported that some members of the cadherin superfamily are involved ingeneral cell-cell interaction processes including transduction. See,Suzuki, J. Cell Biochem., 61(4):531-542 (1996). Therefore, novel membersof the cadherin superfamily are of interest.

[0185] More generally, all novel proteins are of interest, includingmembrane-bound proteins. Membrane- bound proteins and receptors can playan important role in the formation, differentiation and maintenance ofmulticellular organisms. The fate of many individual cells, e.g.,proliferation, migration, differentiation, or interaction with othercells, is typically governed by information received from other cellsand/or the immediate environment. This information is often transmittedby secreted polypeptides (for instance, mitogenic factors, survivalfactors, cytotoxic factors, differentiation factors, neuropeptides, andhormones) which are, in turn, received and interpreted by diverse cellreceptors or membrane-bound proteins. Such membrane-bound proteins andcell receptors include, but are not limited to, cytokine receptors,receptor kinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor.

[0186] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0187] Efforts are being undertaken by both industry and academia toidentify new, native secreted and membrane-bound receptor proteins,particularly membrane bound proteins having identity with cadherins. Theresults of such efforts are provided herein.

[0188] 60. PRO322

[0189] Proteases are enzymatic proteins which are involved in a largenumber of very important biological processes in mammalian andnon-mammalian organisms. Numerous different protease enzymes from avariety of different mammalian and non-mammalian organisms have beenboth identified and characterized, including the serine proteases whichexhibit specific activity toward various serine-containing proteins. Themammalian protease enzymes play important roles in biological processessuch as, for example, protein digestion, activation, inactivation, ormodulation of peptide hormone activity, and alteration of the physicalproperties of proteins and enzymes.

[0190] Neuropsin is a novel serine protease whose mRNA is expressed inthe central nervous system. Mouse neuropsin has been cloned, and studieshave shown that it is involved in the hippocampal plasticity. Neuropsinhas also been indicated as associated with extracellular matrixmodifications and cell migrations. See, generally, Chen, et al.,Neurosci., 7(2):5088-5097 (1995) and Chen, et al., J. Histochem.Cytochem., 46:313-320 (1998).

[0191] Efforts are being undertaken by both industry and academia toidentify new, native membrane-bound or secreted proteins, particularlythose having homology to neuropsin, serine protease, neurosin andtrypsinogen. Many efforts are focused on the screening of mammalianrecombinant DNA libraries to identify the coding sequences for novelsecreted and membrane-bound receptor proteins. Examples of screeningmethods and techniques are described in the literature [see, forexample, Klein et al., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S.Pat. No. 5,536,637)].

[0192] 61. PRO526

[0193] Protein-protein interactions include those involved with receptorand antigen complexes and signaling mechanisms. As more is known aboutthe structural and functional mechanisms underlying protein-proteininteractions, protein-protein interactions can be more easilymanipulated to regulate the particular result of the protein-proteininteraction. Thus, the underlying mechanisms of protein-proteininteractions are of interest to the scientific and medical community.

[0194] All proteins containing leucine-rich repeats are thought to beinvolved in protein-protein interactions. Leucine-rich repeats are shortsequence motifs present in a number of proteins with diverse functionsand cellular locations. The crystal structure of ribonuclease inhibitorprotein has revealed that leucine-rich repeats correspond to beta-alphastructural units. These units are arranged so that they form a parallelbeta-sheet with one surface exposed to solvent, so that the proteinacquires an unusual, nonglobular shape. These two features have beenindicated as responsible for the protein-binding functions of proteinscontaining leucine-rich repeats. See, Kobe and Deisenhofer, TrendsBiochem. Sci., 19(10):415-421 (October 1994).

[0195] A study has been reported on leucine-rich proteoglycans whichserve as tissue organizers, orienting and ordering collagen fibrilsduring ontogeny and are involved in pathological processes such as woundhealing, tissue repair, and tumor stroma formation. lozzo, R. V., Crit.Rev. Biochem. Mol. Biol., 32(2):141-174 (1997). Others studiesimplicating leucine rich proteins in wound healing and tissue repair areDe La Salle, C., et al., Vouv. Rev. Fr. Hematol. (Germany),37(4):215-222 (1995), reporting mutations in the leucine rich motif in acomplex associated with the bleeding disorder Bernard-Soulier syndrome,Chlemetson, K. J., Thromb. Haemost. (Germany), 74(1):111-116 (July1995), reporting that platelets have leucine rich repeats and Ruoslahti,E. I., et al., WO9110727-A by La Jolla Cancer Research Foundationreporting that decorin binding to transforming growth factorβ hasinvolvement in a treatment for cancer, wound healing and scarring.Related by function to this group of proteins is the insulin like growthfactor (IGF), in that it is useful in wound-healing and associatedtherapies concerned with re-growth of tissue, such as connective tissue,skin and bone; in promoting body growth in humans and animals; and instimulating other growth-related processes. The acid labile subunit(ALS) of IGF is also of interest in that it increases the half-life ofIGF and is part of the IGF complex in vivo. ALS is further described inLeong and Baxter, Mol. Endocrinol., 6(6):870-876 (1992); Baxter, J.Biol. Chem., 264(20):11843-11848 (1989); and Khosravi, et al., J. Clin.Endocrinol. Metab., 82(12):3944-3951 (1997).

[0196] Another protein which has been reported to have leucine-richrepeats is the SLIT protein which has been reported to be useful intreating neuro-degenerative diseases such as Alzheimer's disease, nervedamage such as in Parkinson's disease, and for diagnosis of cancer, see,Artavanistsakonas, S. and Rothberg, J. M., WO9210518-A1 by YaleUniversity. Also of interest is LIG-1, a membrane glycoprotein that isexpressed specifically in glial cells in the mouse brain, and hasleucine rich repeats and immunoglobulin-like domains. Suzuki, et al., J.Biol. Chem. (U.S.), 271(37):22522 (1996). Other studies reporting on thebiological functions of proteins having leucine rich repeats include:Tayar, N., et al., Mol. Cell Endocrinol., (Ireland), 125(1-2):65-70(December 1996) (gonadotropin receptor involvement); Miura, Y., et al.,Nippon Rinsho (Japan), 54(7):1784-1789 (July 1996) (apoptosisinvolvement); Harris, P. C., et al., J. Am. Soc. Nephrol.,6(4):1125-1133 (October 1995) (kidney disease involvement).

[0197] Efforts are therefore being undertaken by both industry andacademia to identify new proteins having leucine rich repeats to betterunderstand protein-protein interactions. Of particular interest arethose proteins having leucine rich repeats and identity or similarity toknown proteins having leucine rich repeats such as ALS.

[0198] Many efforts are focused on the screening of mammalianrecombinant DNA libraries to identify the coding sequences for novelsecreted and membrane-bound proteins having leucine rich repeats.Examples of screening methods and techniques are described in theliterature [see, for example, Klein et al., Proc. Natl. Acad. Sci.,93:7108-7113 (1996); U.S. Pat. No. 5,536,637)].

[0199] 62. PRO531

[0200] Cadherins are a large family of transmembrane proteins. Cadherinscomprise a family of calcium-dependent glycoproteins that function inmediating cell-cell adhesion in virtually all solid tissues ofmulticellular organisms. At least cadherins 1-13 as well as types B, E,EP, M, N, P and R have been characterized. Among the functions cadherinsare known for, with some exceptions, cadherins participate in cellaggregation and are associated with cell-cell adhesion sites. Recently,it has been reported that while all cadherins share multiple repeats ofa cadherin specific motif believed to correspond to folding ofextracellular domains, members of the cadherin superfamily havedivergent structures and, possibly, functions. In particular it has beenreported that members of the cadherin superfamily are involved in signaltransduction. See, Suzuki, J. Cell Biochem., 61(4):531-542 (1996).Cadherins are further described in Tanihara, et al., J. Cell Sci.,107(6):1697-1704 (1994), Aberle, et al., J. Cell Biochem., 61(4):514-523(1996) and Tanihara, et al., Cell Adhes. Commun., 2(1):15-26 (1994).

[0201] Protocadherins are members of the cadherin superfamily which arehighly expressed in the brain. In some studies, protocadherins haveshown cell adhesion activity. See, Sano, et al., EMBO J.,12(6):2249-2256 (1993). However, studies have also shown that someprotocadherins, such as protocadherin 3 (also referred to as Pcdh3 orpc3), do not show strong calcium dependent cell aggregation activity.See, Sago, et al., Genomics, 29(3):631-640 (1995) for this study andfurther characteristics of Pcdh3.

[0202] Therefore, novel members of the cadherin superfamily are ofinterest. More generally, all membrane-bound proteins and receptors areof interest. Such proteins can play an important role in the formation,differentiation and maintenance of multicellular organisms. The fate ofmany individual cells, e.g., proliferation, migration, differentiation,or interaction with other cells, is typically governed by informationreceived from other cells and/or the immediate environment. Thisinformation is often transmitted by secreted polypeptides (for instance,mitogenic factors, survival factors, cytotoxic factors, differentiationfactors, neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Suchmembrane-bound proteins and cell receptors include, but are not limitedto, cytokine receptors, receptor kinases, receptor phosphatases,receptors involved in cell-cell interactions, and cellular adhesinmolecules like selectins and integrins. For instance, transduction ofsignals that regulate cell growth and differentiation is regulated inpart by phosphorylation of various cellular proteins. Protein tyrosinekinases, enzymes that catalyze that process, can also act as growthfactor receptors. Examples include fibroblast growth factor receptor andnerve growth factor receptor.

[0203] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inlubitors of the relevant receptor/ligandinteraction.

[0204] Efforts are therefore being undertaken by both industry andacademia to identify new, native membrane bound proteins, particularthose having sequence identity with protocadherins, especially 3 and 4.Many efforts are focused on the screening of mammalian recombinant DNAlibraries to identify the coding sequences for novel membrane-boundproteins. Provided herein are the results of such efforts.

[0205] 63. PRO534

[0206] Protein disulfide isomerase is an enzymatic protein which isinvolved in the promotion of correct refolding of proteins through theestablishment of correct disulfide bond formation. Protein disulfideisomerase was initially identified based upon its ability to catalyzethe renaturation of reduced denatured RNAse (Goldberger et al., J. Biol.Chem. 239:1406-1410 (1964) and Epstein et al., Cold Spring Harbor Symp.Quant. Biol. 28:439-449 (1963)). Protein disulfide isomerase has beenshown to be a resident enzyme of the endoplasmic reticulum which isretained in the endoplasmic reticulum via a -KDEL or -HDEL amino acidsequence at its C-terminus. Protein disulfide isomerase and relatedproteins are further described in Laboissiere, et al., J. Biol. Chem.,270(47:28006-28009 (1995); Jeenes, et al., Gene, 193(2):151-156 (1997;Koivunen, et al., Genomics, 42(3):397-404 (1997); and Desilva, et al.,DNA Cell Biol., 15(1):9-16 (1996). These studies indicate the importanceof the identification of protein disulfide related proteins.

[0207] More generally, and also of interest are all novel membrane-boundproteins and receptors. Such proteins can play an important role in theformation, differentiation and maintenance of multicellular organisms.The fate of many individual cells, e.g., proliferation, migration,differentiation, or interaction with other cells, is typically governedby information received from other cells and/or the immediateenvironment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor.

[0208] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0209] Given the importance of membrane bound proteins, efforts areunder way to identity novel membrane bound proteins. Moreover, given theimportance of disulfide bond-forming enzymes and their potential uses ina number of different applications, for example in increasing the yieldof correct refolding of recombinantly produced proteins, efforts arecurrently being undertaken by both industry and academia to identifynew, native proteins having sequence identity with protein disulfideisomerase. Many of these efforts are focused on the screening ofmammalian recombinant DNA libraries to identify the coding sequences fornovel protein disulfide isomerase homologs. We herein describe a novelpolypeptide having sequence identity with protein disulfide isomeraseand the nucleic acids encoding the same.

[0210] 64. PRO697

[0211] Secreted frizzled related proteins (sFRPs) are related to thefrizzled family of transmembrane receptors. The sFRPs are approximately30 kDa in size, and each contains a putative signal sequence, afrizzled-like cysteine-rich domain, and a conserved hydrophiliccarboxy-terminal domain. It has been reported that sFRPs may function tomodulate Wnt signaling, or function as ligands for certain receptors.Rattner, et al., PNAS USA, 94(7):2859-2863 (1997). Therefore, sFRPs andproteins having sequence identity and/or similarity to sFRPs are ofinterest.

[0212] Another secreted protein of interest is any member of the familyof secreted apoptosis-related proteins (SARPs). Expression of SARPsmodifies the intracellular levels of beta-catenin, suggesting that SARPsinterfere with the Wnt-frizzled proteins signaling pathway. Melkonyan,et al., PNAS USA, 94(25):13636-13641 (1997). Therefore, SARPs andproteins having sequence identity and/or similarity to SARPs are ofinterest.

[0213] In addition to sFRPs and SARPs, many extracellular proteins areof interest. Extracellular proteins play an important role in theformation, differentiation and maintenance of multicellular organisms.The fate of many individual cells, e.g., proliferation, migration,differentiation, or interaction with other cells, is typically governedby information received from other cells and/or the immediateenvironment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. These secreted polypeptides or signalingmolecules normally pass through the cellular secretory pathway to reachtheir site of action in the extracellular environment.

[0214] Secreted proteins have various industrial applications, includingpharmaceuticals, diagnostics, biosensors and bioreactors. Most proteindrugs available at present, such as thrombolytic agents, interferons,interleukins, erythropoietins, colony stimulating factors, and variousother cytokines, are secretory proteins. Their receptors, which aremembrane proteins, also have potential as therapeutic or diagnosticagents.

[0215] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins, particularly those havingsequence identity or similarity with sFRP-2 and SARP-1. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. Examples ofscreening methods and techniques are described in the literature [see,for example, Klein et al., Proc. Natl. Acad. Sci., 93:7108-7113 (1996);U.S. Pat. No. 5,536,637)].

[0216] 65. PRO717

[0217] Efforts are being undertaken by both industry and academia toidentify new, native transmembrane receptor proteins. Many efforts arefocused on the screening of mammalan recombinant DNA libraries toidentify the coding sequences for novel receptor proteins. The resultsof such efforts are provided herein.

[0218] 66. PRO731

[0219] Cadherins are a large family of transmembrane proteins. Cadherinscomprise a family of calcium-dependent glycoproteins that function inmediating cell-cell adhesion in virtally all solid tissues ofmulticellular organisms. At least cadherins 1-13 as well as types B, E,EP, M, N, P and R have been characterized. Among the functions cadherinsare known for, with some exceptions, cadherins participate in cellaggregation and are associated with cell-cell adhesion sites. Recently,it has been reported that while all cadherins share multiple repeats ofa cadherin specific motif believed to correspond to folding ofextracellular domains, members of the cadherin superfamily havedivergent structures and, possibly, functions. In particular it has beenreported that members of the cadherin superfamily are involved in signaltransduction. See, Suzuki, J. Cell Biochem., 61(4):531-542 (1996).Cadherins are further described in Tanihara, et al., J. Cell Sci.,107(6):1697-1704 (1994), Aberle, et al., J. Cell Biochem., 61(4):514-523(1996) and Tanihara, et al., Cell Adhes. Commun., 2(1):15-26 (1994).

[0220] Protocadherins are members of the cadherin superfamily which arehighly expressed in the brain. In some studies, protocadherins haveshown cell adhesion activity. See, Sano, et al., EMBO J.,12(6):2249-2256 (1993). However, studies have also shown that someprotocadherins, such as protocadherin 3 (also referred to as Pcdh3 orpc3), do not show strong calcium dependent cell aggregation activity.See, Sago, et al., Genomics, 29(3):631-640 (1995) for this study andfurther characteristics of Pcdh3.

[0221] Therefore, novel members of the cadherin superfamily are ofinterest. More generally, all membrane- bound proteins and receptors areof interest. Such proteins can play an important role in the formation,differentiation and maintenance of multicellular organisms. The fate ofmany individual cells, e.g., proliferation, migration, differentiation,or interaction with other cells, is typically governed by informationreceived from other cells and/or the immediate environment. Thisinformation is often transmitted by secreted polypeptides (for instance,mitogenic factors, survival factors, cytotoxic factors, differentiationfactors, neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Suchmembrane-bound proteins and cell receptors include, but are not limitedto, cytokine receptors, receptor kinases, receptor phosphatases,receptors involved in cell-cell interactions, and cellular adhesinmolecules like selectins and integrins. For instance, transduction ofsignals that regulate cell growth and differentiation is regulated inpart by phosphorylation of various cellular proteins. Protein tyrosineInases, enzymes that catalyze that process, can also act as growthfactor receptors. Examples include fibroblast growth factor receptor andnerve growth factor receptor.

[0222] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0223] Efforts are therefore being undertaken by both industry andacademia to identify new, native membrane bound proteins, particularthose having sequence identity with protocadherins, especially 4, 68,43, 42, 3 and 5. Many efforts are focused on the screening of mammalianrecombinant DNA libraries to identify the coding sequences for novelmembrane-bound proteins. Provided herein are the results of suchefforts.

[0224] 67. PRO218

[0225] Efforts are being undertaken by both industry and academia toidentify new, native membrane bound proteins, particularly those havingsequence identity with membrane regulator proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel receptor proteins.

[0226] 68. PRO768

[0227] The integrins comprise a supergene family of cell-surfaceglycoprotein receptors that promote cellular adhesion. Each cell hasnumerous receptors that define its cell adhesive capabilities. Integrinsare involved in a wide variety of interaction between cells and othercells or matrix components. The integrins are of particular importancein regulating movement and function of immune system cells. The plateletIIb/IIIA integrin complex , is of particular importance in regulatingplatelet aggregation. A member of the integrin family, integrin P-6, isexpressed on epithelial cells and modulates epithelial inflammation.Another integrin, leucocyte-associated antigen-1 (LFA-1) is important inthe adhesion of lymphocytes during an immune response.

[0228] Of particular interest is H36alpha 7, an integrin alpha chainthat is developmentally regulated during myogenesis as described inSong, et al., J. Cell Biol., 117(3):643-657 (1992). The expressionpattern of the laminin-binding alpha 7 beta 1 integrin isdevelopmentally regulated in skeletal, cardiac, and smooth muscle.Ziober, et al., Mol. Biol. Cell, 8(9):1723-1734 (1997). It has beenreported that expression of the alpha 7-X1/X2 integrin is a novelmechanism that regulates receptor affinity states in a cell-specificcontext and may modulate integrin-dependent events during muscledevelopment and repair. Id. It has further been reported that lamininspromote the locomotion of skeletal myoblasts via the alpha 7 integrinreceptor. In particular it was reported that alpha 7 beta 1 receptor canpromote myoblast adhesion and motility on a restricted number of lamininisoforms and may be important in myogenic precursor recruitment duringregeneration and differentiation. Yao, et al., J. Cell Sci.,109(13):3139-3150 (1996). Spliced variants of integrin alpha 7 are alsodescribed in Leung, et al., Biochem. Biophys. Res. Commun.,243(1):317-325 (1998) and Fornaro and Languino, Matrix Biol.,16(4):185-193 (1997). Moreover, it has been reported that absence ofintegrin alpha 7 causes a form of muscular dystrophy. Thus integrins,particularly those related to integrin 7 and related molecules, are ofinterest.

[0229] In addition to the interest of integrins, more generally, allmembrane-bound proteins and receptors are of interest since suchproteins can play an important role in the formation, differentiationand maintenance of multicellular organisms. The fate of many individualcells, e.g., proliferation, migration, differentiation, or interactionwith other cells, is typically governed by information received fromother cells and/or the immediate environment. This information is oftentransmitted by secreted polypeptides (for instance, mitogenic factors,survival factors, cytotoxic factors, differentiation factors,neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Suchmembrane-bound proteins and cell receptors include, but are not limitedto, cytokine receptors, receptor kinases, receptor phosphatases,receptors involved in cell-cell interactions, and cellular adhesinmolecules like selectins and integrins. For instance, transduction ofsignals that regulate cell growth and differentiation is regulated inpart by phosphorylation of various cellular proteins. Protein tyrosinekinases, enzymes that catalyze that process, can also act as growthfactor receptors. Examples include fibroblast growth factor receptor andnerve growth factor receptor.

[0230] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0231] Therefore, efforts are being undertaken by both industry andacademia to identify new, native receptor proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel receptor proteins. The resultsof such efforts, particularly those focused on identifying newpolypeptides having sequence identity with integrins, are providedherein.

[0232] 69. PRO771

[0233] Testican is a multidomain testicular proteoglycan which isexpressed in numerous tissue types including, but not limited toneuromuscular tissue, the brain and reproductive tissues. Testicanresembles modulators of cell social behavior such as the regulation ofcell shape, adhesion, migration and proliferation. [Bonnet, F. et al.,J. Biol. Chem., 271(8):4373 (1996), Perin, J. P. et al., EXS(Switzerland), 70:191 (1994), Alliel, P. M., et al, Eur. J. Biochem.,214(1):346 (1993), Charbonnier, F., et al., C. R. Seances Soc. Biol.Fil. (France), 191(1):127 (1997)]. Among other reasons, since testicanhas been implicated in neuronal processes and may be associated with thegrowth of connective tissue, testican and related molecules are ofinterest.

[0234] More generally, all extracellular proteins are of interest.Extracellular proteins play an important role in the formation,differentiation and maintenance of multicellular organisms. The fate ofmany individual cells, e.g., proliferation, migration, differentiation,or interaction with other cells, is typically governed by informationreceived from other cells and/or the immediate environment. Thisinformation is often transmitted by secreted polypeptides (for instance,mitogenic factors, survival factors, cytotoxic factors, differentiationfactors, neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Thesesecreted polypeptides or signaling molecules normally pass through thecellular secretory pathway to reach their site of action in theextracellular environment.

[0235] Secreted proteins have various industrial applications, includingpharmaceuticals, diagnostics, biosensors and bioreactors. Most proteindrugs available at present, such as thrombolytic agents, interferons,interleukins, erythropoietins, colony stimulating factors, and variousother cytokines, are secretory proteins. Their receptors, which aremembrane proteins, also have potential as therapeutic or diagnosticagents. Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. Examples of screening methods andtechniques are described in the literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)]. The results of such efforts, particularly those focused onidentifying molecules having identity and/or similarity with testicanare of interest.

[0236] 70. PRO733

[0237] T1/ST2 is a receptor-like molecule homologous to the type Iinterleukin-1 receptor, believed to be involved in cell signaling. TheT1/ST2 receptor and/or putative ligands are further described in Gayle,et al., J. Biol. Chem., 271(10):5784-5789 (1996), Kumar, et al., J.Biol. Chem., 270(46):27905-27913 (1995), and Mitcham, et al., J. Biol.Chem., 271(10):5777-5783 (1996). These proteins, and proteins relatedthereto are of interest.

[0238] More generally all membrane-bound proteins and receptors are ofinterest since they can play an important role in the formation,differentiation and maintenance of multicellular organisms. The fate ofmany individual cells, e.g., proliferation, migration, differentiation,or interaction with other cells, is typically governed by informationreceived from other cells and/or the immediate environment. Thisinformation is often transmitted by secreted polypeptides (for instance,mitogenic factors, survival factors, cytotoxic factors, differentiationfactors, neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Suchmembrane-bound proteins and cell receptors include, but are not limitedto, cytokine receptors, receptor kinases, receptor phosphatases,receptors involved in cell-cell interactions, and cellular adhesinmolecules like selectins and integrins. For instance, transduction ofsignals that regulate cell growth and differentiation is regulated inpart by phosphorylation of various cellular proteins. Protein tyrosinekinases, enzymes that catalyze that process, can also act as growthfactor receptors. Examples include fibroblast growth factor receptor andnerve growth factor receptor.

[0239] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0240] Efforts are being undertaken by both industry and academia toidentify new, native receptor proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel receptor proteins. The results of such efforts areprovided herein.

[0241] 71. PRO162

[0242] Pancreatitis-associated protein (PAP) is a secretory protein thatis overexpressed by the pancreas during acute pancreatitis. Serum PAPconcentrations have been shown to be abnormally high in patients withacute pancreatitis. Pezzirli et al., Am. J. Gastroenterol.,92(10):1887-1890 (1997).

[0243] PAP is synthesized by the pancreas due to pancreatic inflammationand has been shown to be a good serum marker for injury of the pancreas.In addition, serum PAP levels appear to strongly correlate withcreatinine clearance measurements. In patients with a pancreas-kidneytransplantation, PAP may prove to be a useful biological andhistological marker of pancreatic graft rejection. Van der Pijl et al.,Transplantation, 63(7):995-1003 (1997). Further, PAP has been shown tobe useful in screening neonates for cystic fibrosis. In fact, PAP maydiscriminate cystic fibrosis neonates with better specificity than thecurrent immunoreactive trypsis assay. Iovanna et al., C. R. Acad. Aci.III, 317(6):561-564.

[0244] Secreted proteins such as PAP have various industrialapplications, including pharmaceuticals, diagnostics, biosensors andbioreactors. Most protein drugs available at present, such asthrombolytic agents, interferons, interleukins, erythropoietins, colonystimulating factors, and various other cytokines, are secretoryproteins. Their receptors, which are membrane proteins, also havepotential as therapeutic or diagnostic agents.

[0245] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. Examples of screening methods andtechniques are described in the literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)]. The results of such efforts are presented herein.

[0246] 72. PRO788

[0247] Anti-neoplastic urinary protein (ANUP) was identified as themajor protein present in a fraction of human urine which exhibitsantiproliferative activity against human tumor cell lines withoutaffecting the growth of several normal diploid cell lines or tumor cellsof mouse or hamster origin. Sloane et al., Biochem. J., 234(2):355-362(1986).

[0248] ANUP is a unique cytokine that has been found in humangranulocytes. The N-terminal amino acid sequence has been shown to beunique. A synthetic peptide corresponding to the first nine residues,with Cys at positions 4 and 7, was found to be an anti-tumor agent invitro. Ridge and Sloane, Cytokine, 8(1):1-5 (1996).

[0249] Secreted proteins such as ANUP have various industrialapplications, including pharmaceuticals, diagnostics, biosensors andbioreactors. Most protein drugs available at present, such asthrombolytic agents, interferons, interleukins, erythropoietins, colonystimulating factors, and various other cytokines, are secretoryproteins. Their receptors, which are membrane proteins, also havepotential as therapeutic or diagnostic agents. Efforts are beingundertaken by both industry and academia to identify new, nativesecreted proteins. Many efforts are focused on the screening ofmammalian recombinant DNA libraries to identify the coding sequences fornovel secreted proteins. Examples of screening methods and techniquesare described in the literature [see, for example, Klein et al., Proc.Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No. 5,536,637)].

[0250] 73. PRO1008

[0251] Dickkopf-1 (dkk-1) is a member of a family of secreted proteinsand functions in head induction. Dkk-1 is an inducer of Spemannorganizer in amphibian embryos. Glinka, et al., Nature,391(6665):357-362 (1998). Dkk-1 is a potent antagonist of Wntsignalling, suggesting that dkk genes encode a family of secreted Wntinhibitors. Thus, dkk-1 family members and related molecules are ofinterest.

[0252] More generally, all extracellular proteins are of interest sincethey can play an important role in the formation, differentiation andmaintenance of multicellular organisms. The fate of many individualcells, e.g., proliferation, migration, differentiation, or interactionwith other cells, is typically governed by information received fromother cells and/or the immediate environment. This information is oftentransmitted by secreted polypeptides (for instance, mitogenic factors,survival factors, cytotoxic factors, differentiation factors,neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Thesesecreted polypeptides or signaling molecules normally pass through thecellular secretory pathway to reach their site of action in theextracellular environment.

[0253] Secreted proteins have various industrial applications, includingpharmaceuticals, diagnostics, biosensors and bioreactors. Most proteindrugs available at present, such as thrombolytic agents, interferons,interleukins, erythropoietins, colony stimulating factors, and variousother cytokines, are secretory proteins. Their receptors, which aremembrane proteins, also have potential as therapeutic or diagnosticagents.

[0254] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins, particularly those related todkk-1. Many efforts are focused on the screening of mammalianrecombinant DNA libraries to identify the coding sequences for novelsecreted proteins. Examples of screening methods and techniques aredescribed in the literature [see, for example, Klein et al., Proc. Natl.Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No. 5,536,637)]. The resultsof such efforts to identify molecules related to dkk-1 are providedherein.

[0255] 74. PRO1012

[0256] Protein disulfide isomerase is an enzymatic protein which isinvolved in the promotion of correct refolding of proteins through theestablishment of correct disulfide bond formation. Protein disulfideisomerase was initially identified based upon its ability to catalyzethe renaturation of reduced denatured RNAse (Goldberger et al., J. Biol.Chem. 239:1406-1410 (1964) and Epstein et al., Cold Spring Harbor Symp.Quant. Biol. 28:439-449 (1963)). Protein disulfide isomerase has beenshown to be a resident enzyme of the endoplasmic reticulum which isretained in the endoplasmic reticulum via a -KDEL or -HDEL amino acidsequence at its C-terminus. Protein disulfide isomerase and relatedproteins are further described in Laboissiere, et al., J. Biol. Chem.,270(47:28006-28009 (1995); Jeenes, et al., Gene, 193(2):151-156 (1997;Koivunen, et al., Genomics, 42(3):397-404 (1997); and Desilva, et al.,DNA Cell Biol., 15(1):9-16 (1996). These studies indicate the importanceof the identification of protein disulfide related proteins.

[0257] More generally, the identification of all extracellular andmembrane-bound proteins is of interest since they play important rolesin the formation, differentiation and maintenance of multicellularorganisms. The fate of many individual cells, e.g., proliferation,migration, differentiation, or interaction with other cells, istypically governed by information received from other cells and/or theimmediate environment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. These secreted polypeptides or signalingmolecules normally pass through the cellular secretory pathway to reachtheir site of action in the extracellular environment, usually at amembrane-bound receptor protein.

[0258] Secreted proteins have various industrial applications, includinguse as pharmaceuticals, diagnostics, biosensors and bioreactors. Infact, most protein drugs available at present, such as thrombolyticagents, interferons, interleukins, erythropoietins, colony stimulatingfactors, and various other cytokines, are secretory proteins. Theirreceptors, which are membrane-bound proteins, also have potential astherapeutic or diagnostic agents. Receptor immunoadhesins, for instance,can be employed as therapeutic agents to block receptor-ligandinteraction. Membrane-bound proteins can also be employed for screeningof potential peptide or small molecule inhibitors of the relevantreceptor/ligand interaction. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cellHellinteractions, and cellular adhesin molecules like selectins andintegrins. Transduction of signals that regulate cell growth anddifferentiation is regulated in part by phosphorylation of variouscellular proteins. Protein tyrosine kinases, enzymes that catalyze thatprocess, can also act as growth factor receptors. Examples includefibroblast growth factor receptor and nerve growth factor receptor.

[0259] Of particular interest are cellular proteins having endoplasmicreticulum (ER) retention signals. These proteins are retained in thecell and function closely with endoplasmic reticulum in proteinproduction. Such proteins have been described previously, i.e., seeShorrosh and Dixon, Plant J., 2(1):51-58 (1992).

[0260] Efforts are being undertaken by both industry and academia toidentif new, native secreted and membrane-bound receptor proteins, andin particular, cellular proteins having ER retension signals. Manyefforts are focused on the screening of mammalian recombinant DNAlibraries to identify the coding sequences for novel secreted andmembrane-bound receptor proteins. Examples of screening methods andtechniques are described inthe literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)]. The results of such efforts, particularly theidentification of novel polypeptides and nucleic acids encoding thesame, which have sequence identity and similarity to protein disulfideisomerase are presented herein.

[0261] 75. PRO1014

[0262] Oxygen free radicals and antioxidants appear to play an importantrole in the central nervous system after cerebral ischemia andreperfision. Moreover, cardiac injury, related to ischaemia andreperfusion has been reported to be caused by the action of freeradicals. Additionally, studies have reported that the redox state ofthe cell is a pivotal determinant of the fate of the cells. Furthermore,reactive oxygen species have been reported to be cytotoxic, causinginflammatory disease, including tissue necrosis, organ failure,atherosclerosis, infertility, birth defects, premature aging, mutationsand malignancy. Thus, the control of oxidation and reduction isimportant for a number of reasons including for control and preventionof strokes, heart attacks, oxidative stress and hypertension. In thisregard, reductases, and particularly, oxidoreductases, are of interest.Publications further describing this subject matter include Kelsey, etal., Br. J. Cancer, 76(7):8524 (1997); Friedrich and Weiss, J. Theor.Biol., 187(4):52940 (1997) and Pieulle, et al., J. Bacteriol.,179(18):5684-92 (1997).

[0263] In addition to reductases in particular, novel polypeptides aregenerally of interest. Extracellular proteins play an important role inthe formation, differentiation and maintenance of multicellularorganisms. The fate of many individual cells, e.g., proliferation,migration, differentiation, or interaction with other cells, istypically governed by information received from other cells andlor theimmediate environment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. These secreted polypeptides or signalingmolecules normally pass through the cellular secretory pathway to reachtheir site of action in the extracellular environment.

[0264] Secreted proteins have various industrial applications, includingpharmaceuticals, diagnostics, biosensors and bioreactors. Most proteindrugs available at present, such as thrombolytic agents, interferons,interleukins, erythropoietins, colony stimulating factors, and variousother cytokines, are secretory proteins. Their receptors, which aremembrane proteins, also have potential as therapeutic or diagnosticagents. Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel secreted proteins. Examples of screening methods andtechniques are described in the literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)]. The results of such efforts, particularly those identifyingpolypeptides having sequence identity with reductases, and the nucleicacids encoding the same, are presented herein.

[0265] 76. PRO1017

[0266] Enzymatic proteins play important roles in the chemical reactionsinvolved in the digestion of foods, the biosynthesis of macromolecules,the controlled release and utilization of chemical energy, and otherprocesses necessary to sustain life. Sulfotransferases are enzymes whichtransfer sulfate from a sulfate donor to acceptor substrates,particularly those containing terminal glucoronic acid. The HNK-1carbohydrate epitope is expressed on several neural adhesionglycoproteins and a glycolipid, and is involved in cell interactions.The glucuronyltransferase and sulfotransferase are considered to be thekey enzymes in the biosynthesis of this epitope because the rest of thestructure occurs often in glycoconjugates. HNK-1 sulfotransfererase isfurther described in Bakker, H., et al., J. Biol. Chem.,272(47):29942-29946 (1997).

[0267] In addition to HNK-1 sulfotransfererase, and novel proteinsrelated thereto, all novel proteins are of interest. Exacellular andmembrane-bound proteins play important roles in the formation,differentiation and maintenance of multicellular organisms. The fate ofmany individual cells, e.g., proliferation, migration, differentiation,or interaction with other cells, is typically governed by informationreceived from other cells and/or the immediate environment. Thisinformation is often transmitted by secreted polypeptides (for instance,mitogenic factors, survival factors, cytotoxic factors, differentiationfactors, neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Thesesecreted polypeptides or signaling molecules normally pass through thecellular secretory pathway to reach their site of action in theextracellular environment, usually at a membrane-bound receptor protein.

[0268] Secreted proteins have various industrial applications, includinguse as pharmaceuticals, diagnostics, biosensors and bioreactors. Infact, most protein drugs available at present, such as thrombolyticagents, interferons, interleukins, erythropoietins, colony stimulatingfactors, and various other cytokines, are secretory proteins. Theirreceptors, which are membrane-bound proteins, also have potential astherapeutic or diagnostic agents. Receptor immunoadhesins, for instance,can be employed as therapeutic agents to block receptor-ligandinteraction. Membrane-bound proteins can also be employed for screeningof potential peptide or small molecule inhibitors of the relevantreceptor/ligand interaction. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. Transduction of signals that regulate cell growth anddifferentiation is regulated in part by phosphorylation of variouscellular proteins. Protein tyrosine kinases, enzymes that catalyze thatprocess, can also act as growth factor receptors. Examples includefibroblast growth factor receptor and nerve growth factor receptor.

[0269] Efforts are being undertaken by both industry and academia toidentify new, native secreted and membrane-bound receptor proteins,particularly those having sequence identity with HNK-1 sulfotransferase.Many efforts are focused on the screening of mammalian recombinant DNAlibraries to identify the coding sequences for novel secreted andmembrane-bound receptor proteins. Examples of screening methods andtechniques are described in the literature [see, for example, Klein etal., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S. Pat. No.5,536,637)]. The results of such efforts are provided herein.

[0270] 77. PRO474

[0271] Enzymatic proteins play important roles in the chemical reactionsinvolved in the digestion of foods, the biosynthesis of macromolecules,the controlled release and utilization of chemical energy, and otherprocesses necessary to sustain life. Glucose dehydrogenase functions inthe oxidation of glucose to gluconate to generate metabolically usefulenergy. The regulation of the PQQ-linked glucose dehydrogenase indifferent organisms is reviewed in Neijssel, et al., Antonie VanLeeuwenhoek, 56(1):51-61 (1989). Glucose dehydrogenase functions as anauxiliary energy generating mechanism, because it is maximallysynthesized under conditions of energy stress. In addition to moleculesrelated to glucose dehydrogenase, all novel proteins are of interest.Extracellular and membrane-bound proteins play important roles in theformation, differentiation and maintenance of multicellular organisms.The fate of many individual cells, e.g., proliferation, migration,differentiation, or interaction with other cells, is typically governedby information received from other cells and/or the immediateenvironment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. These secreted polypeptides or signalingmolecules normally pass through the cellular secretory pathway to reachtheir site of action in the extracellular environment, usually at amembrane-bound receptor protein.

[0272] Secreted proteins have various industrial applications, includinguse as pharmaceuticals, diagnostics, biosensors and bioreactors. Infact, most protein drugs available at present, such as thrombolyticagents, interferons, interleukins, erythropoietins, colony stimulatingfactors, and various other cytokines, are secretory proteins. Theirreceptors, which are membrane-bound proteins, also have potential astherapeutic or diagnostic agents. Receptor immunoadhesins, for instance,can be employed as therapeutic agents to block receptor-ligandinteraction. Membrane-bound proteins can also be employed for screeningof potential peptide or small molecule inhibitors of the relevantreceptor/ligand interaction. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. Transduction of signals that regulate cell growth anddifferentiation is regulated in part by phosphorylation of variouscellular proteins. Protein tyrosine kinases, enzymes that catalyze thatprocess, can also act as growth factor receptors. Examples includefibroblast growth factor receptor and nerve growth factor receptor.

[0273] Efforts are being undertaken by both industry and academia toidentify new, native secreted and membrane-bound receptor proteins, andparticularly cellular proteins and those related to dehydrogenase oroxidoreductase. Many efforts are focused on the screening of mammalianrecombinant DNA libraries to identify the coding sequences for novelsecreted and membrane-bound receptor proteins. Examples of screeningmethods and techniques are described in the literature [see, forexample, Klein et al., Proc. Natl. Acad. Sci., 93:7108-7113 (1996); U.S.Pat. No. 5,536,637)]. The results of such efforts are presented herein.

[0274] 78. PRO1031

[0275] It has been reported that the cytokine interleukin 17 (IL-17)stimulates epithelial, endothelial, and fibroblastic cells to secretecytokines such as ILL, IL-8, and granulocyte-colony-stimulating factor,as well as prostaglandin E2. Moreover, it has been shown that whencultured in the presence of IL-17, fibroblasts could sustainproliferation of CD34+ preferential maturation into neutrophils. Thus ithas been suggested that EL-17 constitutes an early initiator of the Tcell-dependent inflammatory reaction and/or an element of the cytokinenetwork that bridges the immune system to hematopoiesis. See, Yao, etal., J. Immunol., 155(12):5483-5486 (1995); Fossiez, et al., J. Exp.Med., 183(6):2593-2603 (1996); Kennedy, et al., J. Interferon CytokineRes., 16(8):611-617 (1996). Thus, proteins related to IL-17 are ofinterest.

[0276] More generally, all novel proteins are of interest. Extracellularproteins play an important role in the formation, differentiation andmaintenance of multicellular organisms. The fate of many individualcells, e.g., proliferation, migration, differentiation, or interactionwith other cells, is typically governed by information received fromother cells and/or the immediate environment. This information is oftentransmitted by secreted polypeptides (for instance, mitogenic factors,survival factors, cytotoxic factors, differentiation factors,neuropeptides, and hormones) which are, in turn, received andinterpreted by diverse cell receptors or membrane-bound proteins. Thesesecreted polypeptides or signaling molecules normally pass through thecellular secretory pathway to reach their site of action in theextracellular environment.

[0277] Secreted proteins have various industrial applications, includingpharmaceuticals, diagnostics, biosensors and bioreactors. Most proteindrugs available at present, such as thrombolytic agents, interferons,interleukins, erythropoietins, colony stimulating factors, and variousother cytokines, are secretory proteins. Their receptors, which aremembrane proteins, also have potential as therapeutic or diagnosticagents.

[0278] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins, particularly those related toIL-17. Many efforts are focused on the screening of mamm an recombinantDNA libraries to identify the coding sequences for novel secretedproteins. Examples of screening methods and techniques are described inthe literature [see, for example, Klein et al., Proc. Natl. Acad. Sci.,93:7108-7113 (1996); U.S. Pat. No. 5,536,637)]. The results of suchefforts are presented herein.

[0279] 79. PRO938

[0280] Protein disulfide isomerase is an enzymatic protein which isinvolved in the promotion of correct refolding of proteins through theestablishment of correct disulfide bond formation. Protein disulfideisomerase was initially identified based upon its ability to catalyzethe renaturation of reduced denatured RNAse (Goldberger et al., J. Biol.Chem. 239:1406-1410 (1964) and Epstein et al., Cold Spring Harbor Symp.Quant. Biol. 28:439-449 (1963)). Protein disulfide isomerase has beenshown to be a resident enzyme of the endoplasmic reticulum which isretained in the endoplasmic reticulum via a -KDEL or -HDEL amino acidsequence at its C-terminus. Protein disulfide isomerase and relatedproteins are further described in Laboissiere, et al., J. Biol. Chem.,270(47):28006-28009 (1995); Jeenes, et al., Gene, 193(2:151-156 (1997);Koivunen, et al., Genomics, 42(3):397-404 (1997); Desilva, et al., DNACell Biol., 15(1):9-16 (1996); Freedman, et al. Trends in Biochem. Sci.19:331-336 (1994); Bulleid, N. J. Advances in Prot. Chem. 44:125-50(1993); and Noiva, R., Prot. Exp. and Purification 5:1-13 (1994). Thesestudies indicate the importance of the identification of proteindisulfide related proteins.

[0281] More generally, and also of interest are all novel membrane-boundproteins and receptors. Such proteins can play an important role in theformation, differentiation and maintenance of multicellular organisms.The fate of many individual cells, e.g., proliferation, migration,differentiation, or interaction with other cells, is typically governedby information received from other cells and/or the immediateenvironment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. Such membrane-bound proteins and cellreceptors include, but are not limited to, cytokine receptors, receptorkinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor.

[0282] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0283] Given the importance of membrane bound proteins, efforts areunder way to identity novel membrane bound proteins. Moreover, given theimportance of disulfide bond-forming enzymes and their potential uses ina number of different applications, for example in increasing the yieldof correct refolding of recombinantly produced proteins, efforts arecurrently being undertaken by both industry and academia to identifynew, native proteins having sequence identity with protein disulfideisomerase. Many of these efforts are focused on the screening ofmammalian recombinant DNA libraries to identify the coding sequences fornovel protein disulfide isomerase homologs.

[0284] We herein describe the identification and characterization of anovel polypeptide having homology to protein disulfide isomerase.

[0285] 80. PRO1082

[0286] The low density lipoprotein (LDL) receptor is a membrane-boundprotein that plays a key role in cholesterol homeostasis, mediatingcellular uptake of lipoprotein particles by high affinity binding to itsligands, apolipoprotein (apo) B-100 and apoE. The ligand-binding domainof the LDL receptor contains 7 cysteine-rich repeats of approximately 40amino acids, wherein each repeat contains 6 cysteines, which form 3intra-repeat disulfide bonds. These unique structural features providethe LDL receptor with its ability to specifically interact with apoB-100 and apoE, thereby allowing for transport of these lipoproteinparticles across cellular membranes and metabolism of their components.Soluble fragments containing the extracellular domain of the LDLreceptor have been shown to retain the ability to interact with itsspecific lipoprotein ligands (Simmons et al., J. Biol. Chem.272:25531-25536 (1997)). LDL receptors are further described in Javitt,FASEB J., 9(13):1378-1381 (1995) and Herz and Willnow, Ann. NY Acad.Sci., 737:14-19 (1994). Thus, proteins having sequence identity with LDLreceptors are of interest.

[0287] More generally, all membrane-bound proteins and receptors canplay an important role in the formation, differentiation and maintenanceof multicellular organisms. The fate of many individual cells, e.g.,proliferation, migration, differentiation, or interaction with othercells, is typically governed by information received from other cellsand/or the immediate environment. This information is often transmittedby secreted polypeptides (for instance, mitogenic factors, survivalfactors, cytotoxic factors, differentiation factors, neuropeptides, andhormones) which are, in turn, received and interpreted by diverse cellreceptors or membrane-bound proteins. Such membrane-bound proteins andcell receptors include, but are not limited to, cytokine receptors,receptor kinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor. Of particular interest are membrane bound proteins that havetype II transmembrane domains.

[0288] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0289] Efforts are thus being undertaken by both industry and academiato identify new, native proteins, particularly membrane bound proteinsincluding type 11 transmembrane bound proteins. Many efforts are focusedon the screening of mammalian recombinant DNA libraries to identify thecoding sequences for novel receptor proteins. The results of suchefforts are provided herein.

[0290] 81. PRO1083

[0291] Of particular interest are membrane bound proteins that belong tothe seven transmembrane (7TME) receptor superfamily. Examples of thesereceptors include G-protein coupled receptors such as ion receptors.Another example of a 7TM receptor superfamily member is described inOsterhoff, et al., DNA Cell Biol., 16(4):379-389 (1997).

[0292] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interaction. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0293] Efforts are being undertaken by both industry and academia toidentify new, native receptor proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel receptor proteins. The results of such efforts arepresented herein.

[0294] 82. PRO200

[0295] Polypeptides involved in survival, proliferation and/ordifferentiation of cells are of interest. Polypeptides known to beinvolved in the survival, proliferation and/or differentiation of cellsinclude VEGF and members of the bone morphogenetic protein family.Therefore, novel polypeptides which are related to either VEGF or thebone morphogenetic protein are of interest.

[0296] The heparin-binding endothelial cell-growth factor, VEGF, wasidentified and purified from media conditioned by bovine pituitaryfollicular or folliculo-stellate cells over several years ago. SeeFerrara et al., Biophys. Res. Comm. 161, 851 (1989). VEGF is a naturallyoccurring compound that is produced in follicular or folliculo-stellatecells (FC), a morphologically well characterized population of granularcells. The FC are stellate cells that send cytoplasmic processes betweensecretory cells.

[0297] VEGF is expressed in a variety of tissues as multiple homodimericforms (121, 165, 189 and 206 amino acids per monomer) resulting fromalternative RNA splicing. VEGF₁₂₁ is a soluble mitogen that does notbind heparin; the longer forms of VEGF bind heparin with progressivelyhigher affinity. The heparin-binding forms of VEGF can be cleaved in thecarboxy terminus by plasmin to release (a) diffusible form(s) of VEGF.Amino acid sequencing of the carboxy terminal peptide identified afterplasmin cleavage is Arg₁₁₀-Ala₁₁₁. Amino terminal “core” protein, VEGF(1-110) isolated as ahomodirner, binds neutralizingmonoclonalantibodies(4.6.1 and 2E3) and soluble forms of FMS-like tyrosine kinase (FLT-1),kinase domain region (KDR) and fetal liver kinase (FLK) receptors withsimilar affinity compared to the intact VEGF₁₆₅ homodimer.

[0298] As noted, VEGF contains two domains that are responsiblerespectively for binding to the KDR and FLT-1 receptors. These receptorsexist only on endothelial (vascular) cells. As cells become depleted inoxygen, because of trauma and the like, VEGF production increases insuch cells which then bind to the respective receptors in order tosignal ultimate biological effect. The signal then increases vascularpermeability and the cells divide and expand to form new vascularpathways—vasculogenesis and angiogenesis.

[0299] Thus, VEGF is useful for treating conditions in which a selectedaction on the vascular endothelial cells, in the absence of excessivetissue growth, is important, for example, diabetic ulcers and vascularinjuries resulting from trauma such as subcutaneous wounds. Being avascular (artery and venus) endothelial cell growth factor, VEGFrestores cells that are damaged, a process referred to asvasculogenesis, and stimulates the formulation of new vessels, a processreferred to as angiogenesis.

[0300] VEGF would also find use in the restoration of vasculature aftera myocardial infarct, as well as other uses that can be deduced. In thisregard, inhibitors of VEGF are sometimes desirable, particularly tomitigate processes such as angiogenesis and vasculogenesis in cancerouscells.

[0301] Regarding the bone morphogenetic protein family, members of thisfamily have been reported as being involved in the differentiation ofcartilage and the promotion of vascularization and osteoinduction inpreformed hydroxyapatite. Zou, et al., Genes Dev. (U.S.), 11(17):2191(1997); Levine, et al., Ann. Plast. Surg., 39(2):158 (1997). A number ofrelated bone morphogenetic proteins have been identified, all members ofthe bone moiphogenetic protein (BMP) family. Bone morphogenetic nativeand mutant proteins, nucleic acids encoding therefor, related compoundsincluding receptors, host cells and uses are further described in atleast: U.S. Pat. Nos. 5,670,338; 5,454,419; 5,661,007; 5,637,480;5,631,142; 5,166,058; 5,620,867; 5,543,394; 4,877,864; 5,013,649;55,106,748; and 5,399,677. Of particular interest are proteins havinghomology with bone morphogenetic protein 1, a procollagen C-proteinasethat plays key roles in regulating matrix deposition.

[0302] The present invention is predicated upon research intended toidentify novel polypeptides which are related to VEGF and the BMPfamily, and in particular, polypeptides which have a role in thesurvival, proliferation and/or differentiation of cells. While the novelpolypeptides are not expected to have biological activity identical tothe known polypeptides to which they have homology, the knownpolypeptide biological activities can be used to determine the relativebiological activities of the novel polypeptides. In particular, thenovel polypeptides described herein can be used in assays which areintended to determine the ability of a polypeptide to induce survival,proliferation or differentiation of cells. In turn, the results of theseassays can be used accordingly, for diagnostic and therapeutic purposes.The results of such research is the subject of the present invention.

[0303] 83. PRO285 and PRO286

[0304] The cloning of the Toll gene of Drosophila, a maternal effectgene that plays a central role in the establishment of the embryonicdorsal-ventral pattern, has been reported by Hashiinoto et al., Cell 52,269-279 (1988). The Drosophila Toll gene encodes an integral membraneprotein with an extracytoplasmic domain of 803 amino acids and acytoplasmic domain of 269 amino acids. The extracytoplasmic domain has apotential membrane-spanning segment, and contains multiple copies of aleucine-rich segment, a structural motif found in many transmembraneproteins. The Toll protein controls dorsal-ventral patterning inDrosophila embryos and activates the transcription factor Dorsal uponbinding to its ligand Spätzle. (Morisato and Anderson, Cell 76, 677-688(1994).) In adult Drosophila, the Toll/Dorsal signaling pathwayparticipates in the anti-fungal immune response. (Lenaitre et al., Cell86, 973-983 (1996).)

[0305] A human homologue of the Drosophila Toll protein has beendescribed by Medzhitov et al., Nature 388 394-397 (1997). This humanToll, just as Drosophila Toll, is a type I transmembrane protein, withan extracellular domain consisting of 21 tandemly repeated leucine-richmotifs (teucine-rich region—LRR), separated by a non-LRR region, and acytoplasmic domain homologous to the cytoplasmic domain of the humaninterleukin-1 (IL-1) receptor. A constitutively active mutant of thehuman Toll transfected into human cell lines was shown to be able toinduce the activation of NF-XB and the expression of NF-6controlledgenes for the inflammatory cytokines IL-1, IL-6 and IL-8, as well as theexpression of the constimulatory molecule B7.1, which is required forthe activation of native T cells. It has been suggested that Tollfunctions in vertebrates as a non-clonal receptor of the immune system,which can induce signals for activating both an innate and an adaptiveimmune response in vertebrates. The human Toll gene reported byMedzhitov et al., supra was most strongly expressed in spleen andperipheral blood leukocytes (PBL), and the authors suggested that itsexpression in other tissues may be due to the presence of macrophagesand dendritic cells, in which it could act as an early- warning systemfor infection. The public GenBank database contains the following Tollsequences: Toll (DNAX# HSU88540-1, which is identical with the randomsequenced full-length cDNA #HUMRSC786-1); To112 (DNAX# HSU88878-1);Toll3 (DNAX# HSU88879-1); and Toll4 (DNAX# HSU88880-1, which isidentical with the DNA sequence reported by Medzhitov et al., supra). Apartial Toll sequence (Toll5) is available from GenBank under DNAX#HSU88881-1.

[0306] Further human homologues of the Drosophila Toll protein,designated as Toll-like receptors (huTLRs1-5) were recently cloned andshown to mirror the topographic structure of the Drosophila counterpart(Rock et al., Proc. Natl. Acad. Sci. USA 95, 588-593 [1998]).Overexpression of a constitutively active mutant of one human TLR(Toll-protein homologue—Medzhitov et al., supra; TLR4—Rock et al.,supra) leads to the activation of NF-RκB and induction of theinflammatory cytokines and constimulatory molecules. Medzhitov et al.,supra.

[0307] 84. PRO213-1, PRO1330 and PRO1449

[0308] Cancer is characterized by the increase in the number ofabnormal, or neoplastic, cells derived from a normal tissue whichproliferate to form a tumor mass, the invasion of adjacent tissues bythese neoplastic tumor cells, and the generation of malignant cellswhich eventually spread via the blood or lymphatic system to regionallymph nodes and to distant sites (metastasis). In a cancerous state acell proliferates under conditions in which normal cells would not grow.Cancer manifests itself in a wide variety of forms, characterized bydifferent degrees of invasiveness and aggressiveness.

[0309] Alteration of gene expression is intimately related to theuncontrolled cell growth and de-differentiation which are a commonfeature of all cancers. The genomes of certain well studied tumors havebeen found to show decreased expression of recessive genes, usuallyreferred to as tumor suppression genes, which would normally function toprevent malignant cell growth, and/or overexpression of certain dominantgenes, such as oncogenes, that act to promote malignant growth. Each ofthese genetic changes appears to be responsible for importing some ofthe traits that, in aggregate, represent the full neoplastic phenotype(Hunter, Cell 64, 1129 [1991]; Bishop, Cell 64, 235-248 [1991]).

[0310] A well known mechanism of gene (e.g. oncogene) overexpression incancer cells is gene amplification. This is a process where in thechromosome of the ancestral cell multiple copies of a particular geneare produced. The process involves unscheduled replication of the regionof chromosome comprising the gene, followed by recombination of thereplicated segments back into the chromosome (Alitalo et al., Adv.Cancer Res. 47, 235-281 [1986]). It is believed that the overexpressionof the gene parallels gene amplification, i.e. is proportionate to thenumber of copies made.

[0311] Proto-oncogenes that encode growth factors and growth factorreceptors have been identified to play important roles in thepathogenesis of various human malignancies, including breast cancer. Forexample, it has been found that the human ErbB2 gene (erbB2, also knownas her2, or c-erbB-2), which encodes a 185-kd transmembrane glycoproteinreceptor (p185HER2; HER2) related to the epidermal growth factorreceptor (EGFR), is overexpressed in about 25% to 30% of human breastcancer (Slamon et al., Science 235:177-182 [1987]; Slamon et al.,Science 244:707-712 [1989]).

[0312] It has been reported that gene amplification of a protooncogeneis an event typically involved in the more malignant forms of cancer,and could act as a predictor of clinical outcome (Schwab et al., GenesChromosomes Cancer 1, 181-193 [1990]; Aitalo et al., supra). Thus, erbB2overexpression is commonly regarded as a predictor of a poor prognosis,especially in patients with primary disease that involves axillary lymphnodes (Slamon et al., [1987] and [1989], supra; Ravdin and Chamness,Gene 159:19-27 [1995]; and Hynes and Stem, Biochem Biophys Acta 1198:165-184 [1994]), and has been linked to sensitivity and/or resistance tohormone therapy and chemotherapeutic regimens, including CMF(cyclophosphamide, methotrexate, and fluoruracil) and anthracyclines(Baselga et al., Oncology 11 (3 Suppl 1):4348 [1997]). However, despitethe association of erbB2 overexpression with poor prognosis, the odds ofHER2-positive patients responding clinically to treatment with taxaneswere greater than three times those of HER2-negative patients ([bid). Arecombinant humanized anti-ErbB2 (anti-HER2) monoclonal antibody (ahumanized version of the murine anti-ErbB2 antibody 4D5, referred to asrhuMAb HER2 or Herceptin 7o) has been clinically active in patients withErbB2-overexpressing metastatic breast cancers that had receivedextensive prior anticancer therapy. (Baselga et al., J. Clin. Oncol.14:737-744 [1996]).

[0313] The protein Notch and its homologues are key regulatory receptorsin determining the cell fate in various development processes. Theprotein Notch-4, also known as int-3 oncogene, was originally identifiedas a frequent target in mouse mammary tumor virus (MMVS). Notch-4 isbelieved to be a transgene which affects the differentiation capacity ofstem cells and leads to neoplastic proliferation in epithelial cells.Shirayoshi et al., Genes Cells 2(3):213-224 (1997). Duringembryogenesis, the expression of Notch-4 was detected in endothelialcells of blood vessels forming tissues such as the dorsal aorta,intersegmental vessels, yolk sac vessels, cephalic vessels, heart,vessels in branchial arches, and capillary plexuses. Notch-4 expressionin these tissues was also associated with flk-1, the major regulatorygene of vasculogenesis and angiogenesis. Notch-4 is also upregulated invitro during the differentiation of endothelial stem cell. Theendothelial cell specific expression pattern of Notch-4, as well as itsstructural similarity to Notch suggest that Notch-4 is an endothelialcell specific homologue of Notch and that it may play a role invaculogenesis and angiogenesis.

[0314] 85. PRO298

[0315] Efforts are being undertaken by both industry and academia toidentify new, native receptor proteins. Many efforts are focused on thescreening of mammalian recombinant DNA libraries to identify the codingsequences for novel receptor proteins. We herein describe theidentification and characterization of novel transmembrane polypeptides,designated herein as PRO298 polypeptides.

[0316] 86. PRO337

[0317] Neuronal development in higher vertebrates is characterized byprocesses that must successfully navigate distinct cellular environmenten route to their synaptic targets. The result is a functionally preciseformation of neural circuits. The precision is believed to result formmechanisms that regulate growth cone pathfinding and target recognition,followed by latter refinement and remodeling of such projections byevents that require neuronal activity, Goodman and Shatz, Cell/Neuron[Suppl.] 72(10):77-98 (1993). It is further evident that differentneurons extend nerve fibers that are biochemically distinct and rely onspecific guidance cues provided by cell-cell, cell-matrix, andchemotrophic interactions to reach their appropriate synaptic targets,Goodman et al., supra.

[0318] One particular means by which diversity of the neuronal cellsurface may be generated is through differential expression of cellsurface proteins referred to as cell adhesion molecules (CAMs).Neuronally expressed CAMs have been implicated in diverse developmentalprocesses, including migration of neurons along radial glial cells,providing permissive or repulsive substrates for neurite extension, andin promoting the selective fasciculation of axons in projectionalpathways. lessel, Neuron 1: 3-13 (1988); Edelman and Crossin, Annu. Rev.Biochem. 60: 155-190 (1991). Interactions between CAMs present on thegrowth cone membrane and molecules on opposing cell membranes or in theextracellular matrix are thought to provide the specific guidance cuesthat direct nerve fiber outgrowth along appropriate projectionalpathways. Such interactions are likely to result in the activation ofvarious second messenger systems within the growth cone that regulateneurite outgrowth. Doherty and Walsh, Curr. Opin Neurobiol. 2: 595-601(1992).

[0319] In higher vertebrates, most neural CAMs have been found to bemembers of three major structural families of proteins: the integrins,the cadherins, and the immunoglobulin gene superfamily (IgSF). Jessel,supra.; Takeichi, Annu. Rev. Biochem. 59: 237-252 (1990); Reichardt andTomaselli, Annu. Rev. Neurosci. 14: 531-570 (1991). Cell adhesionmolecules of the IgSF (or Ig-CAMs), in particular, constitute a largefamily of proteins frequently implicated in neural cell interactions andnerve fiber outgrowth during development, Salzer and Colman, Dev.Neurosci. 11: 377-390 (1989); BrWimendorf and Rathjen, J. Neurochem. 61:1207-1219 (1993). However, the majority of mammalian Ig-CAMs appear tobe too widely expressed to specify navigational pathways or synaptictargets suggesting that other CAMs, yet to be identified, have role inthese more selective interactions of neurons.

[0320] Many of the known neural Ig-CAMs have been found to be attachedto the plasma membrane via a glycosylphosphatidylinositol (GPI) anchor.Additionally, many studies have implicated GPI-anchored proteins inproviding specific guidance cues during the outgrowth on neurons inspecific pathways. In studies of the grasshopper nervous system,treatment of embryos with phosphatidylinositol-specific phopholipase C(PIPLC), which selectively removes GPI-anchored proteins from thesurfaces of cells, resulted in misdirection and faulty navigation amongsubsets of pioneering growth cones, as well as inhibited migratorypatterns of a subset of early neurons, Chang et al., Devel. 114: 507-519(1992). The projection of retinal fibers to the optic tectum appears todepend, in part, on a 33 kDa GPI-anchored protein, however, the precisenature of this protein is unknown. Stabl et al., Neuron 5: 735-743(1990).

[0321] The expression of various GPI-anchored proteins has beencharacterized amongst the different populations of primary rat neuronsamongst dorsal root ganglion, sympathetic neurons of the cervicalganglion, sympathetic neurons of the superior cervical ganglion, andcerebellar granule neurons. Rosen et al., J. Cell Biol. 117: 617-627(1992). In contrast to the similar pattern of total membrane proteinexpression by these different types of neurons, striking differenceswere observed in the expression of GPI-anchored proteins between theseneurons. Recently, a 65 kDa protein band known as neurotrimin wasdiscovered and found to be differentially expressed by primary neurons(Rosen et al., supra), and restricted to the nervous system and found tobe the most abundant and earliest expressed of the GPI-anchored speciesin the CNS. Strryk et al., J. Neuroscience 15(3):2141-2156 (1995). Thediscovery of neurotrimin has further lead to the identification of afamily of IgSF members, each containing three Ig-like domains that sharesignificant amino acid identity, now termed IgLON. Struyk et al., supra;Pimenta et al., Gene 170(2):189-95 (1996).

[0322] Additional members of the IgLON subfamily include opiate bindingcell adhesion molecule (OBCAM), Schofield et al., EMBO J. 8: 489-495(1989); limbic associated membrane protein (LAMP), Pimenta et al.,supra; CEPU-1; GP55, Wilson et al., J. Cell Sci. 109: 3129-3138 (1996);Eur. J. Neurosci. 9(2):334-41 (1997); and AvGp50, Hancox et al., BrainRes. Mol. Brain Res. 44(2):273-85 (1997).

[0323] While the expression of neurotrimin appears to be widespread, itdoes appear to correlated with the development of several neuralcircuits. For example, between E18 and P10, neurotimin mRNA expressionwithin the forebrain is maintained at high levels in neurons of thedeveloping thalamus, cortical subplate, and cortex, particularly laminaeV and VI (with less intense expression in II, II, and IV, and minimalexpression in lamina I). Cortical subplate neurons may provide an early,temporary scaffold for the ingrowing thalamic afferents en route totheir final synaptic targets in the cortex. Allendoerfer and Shatz,Annu. Rev. Neurosci. 17: 185-218 (1994). Conversely, subplate neuronshave been suggested to be required for cortical neurons from layer V toselect VI to grow into the thalamus, and neurons from layer V to selecttheir targets in the colliculus, pons, and spinal cord (McConnell etal., J. Neurosci. 14: 1892-1907 (1994). The high level expression ofneurotrimin in many of these projections suggests that it could beinvolved in their development.

[0324] In the hindbrain, high levels of neurotrimin message expressionwere observed within the pontine nucleus and by the internal granulecells and Purkinje cells of the cerebellum. The pontine nucleus receivedafferent input from a variety of sources including corticopontine fibersof layer V, and is a major source of afferent input, via mossy fibers,to the granule cells which, in turn, are a major source of afferentinput via parallel fibers to Purkinje cells. [Palay and Chan-Palay, Thecerebellar cortex: cytology and organization. New York: Springer (1974].High level expression of neurotrimin these neurons again suggestspotential involvement in the establishment of these circuits.

[0325] Neurotrimin also exhibits a graded expression pattern in theearly postnatal striatum. Increased neurotrin expression is foundoverlying the dorsolateral striatum of the rat, while lesserhybridization intensity is seen overlying the ventromedial striatum.Struyk et al., supra. This region of higher neurotrinin hybridizationintensity does not correspond to a cytoarchitecturally differentiableregion, rather it corresponds to the primary area of afferent input fromlayer VI of the contralateral sensorimotor cortex (Gerfen, Nature 311:461-464 (1984); Donoghue and Herkenham, Brain Res. 365: 397-403 (1986)).The ventromedial striatum, by contrast, receives the majority of itsafferent input from the perirhinal and association cortex. It isnoteworthy that a complementary graded pattern of LAMP expression, hasbeen observed within the striatium, with highest expression inventromedial regions, and lowest expression dorsolaterally. Levitt,Science 223: 299-301 (1985); Chesselet et al., Neuroscience 40: 725-733(1991).

[0326] 87. PRO403

[0327] Type II transmembrane proteins, also known as single passtransmembrane proteins have an N-terminal portion lodged in thecytoplasm while the C-ternninal portion is exposed to the extracellulardomain.

[0328] Endothelin is a family of vasoconstrictor peptides about whichmuch activity has been focused to better understand its basicpharmacological, biochemical and molecular biological features,including the presence and structure of isopeptides and their genes(endothelin-1,-2 and û3), regulation of gene expression, intracellularprocessing, specific endothelin converting enzymes (ECE), receptorsubtypes (ET-A and ET-B), intracellular signal transduction followingreceptor activation, etc.

[0329] The endothelin (ET) family of peptides have potent vascular,cardiac and renal actions which may be of pathophysiological importancein many human disease states. ET-1 is expressed as an inactive 212 aminoacid prepropeptide. The prepropeptide is first cleaved at Arg52-Cys53and Arg92-Ala93 and then the carboxy terminal Lys91 and Arg92 aretrimmed from the protein to generate the propeptide big ET-1.

[0330] Endothelin is generated from inactive intermediates, the bigendothelins, by a unique processing event catalyzed by the zincmetalloprotease, endothelin converting enzyme (ECE). ECE was recentlycloned, and its structure was shown to be a single pass transmembraneprotein with a short intracellular N-terminal and a long extracellularC-terminal that contains the catalytic domain and numerousN-glycosylation sites. ECEs cleave the endothelin propeptide betweenTrp73 and Val74 producing the active peptide, ET, which appears tofunction as a local rather than a circulating hormone (Rubanyi, G. M. &Polokoff, M. A., Pharmachological Reviews 46: 325-415 (1994). Thus ECEactivity is a potential site of regulation of endothelin production anda possible target for therapeutic intervention in the endothelin system.By blocking ECE activity, it is possible stop the production of ET-1 byinhibiting the conversion of the relatively inactive precursor, bigET-1, to the physiologically active form.

[0331] Endothelins may play roles in the pathophysiology of a number ofdisease states including: 1) cardiovascular diseases (vasospasm,hypertension, myocardial ischemia; reperfusion injury and acutemyochardial infarction, stroke (cerebral ischemia), congestive heartfailure, shock, atherosclerosis, vascular thickening); 2) kidney disease(acute and chronic renal failure, glomerulonephritis, cirrhosis); 3)lung disease (bronchial asthma, pulmonary hypertension); 4)gastrointestinal disorders (gastric ulcer, inflammatory bowel diseases);5) reproductive disorders (premature labor, dysmenorhea, preeclampsia)and 6) carcinogenesis. Rubanyi & Polokoff, supra.

SUMMARY OF THE INVENTION

[0332] 1. PRO213

[0333] Applicants have identified a cDNA clone that encodes a novelpolypeptide, wherein the polypeptide is designated in the presentapplication as “PRO213”.

[0334] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO213 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO213polypeptide having amino acid residues 1 to 295 of FIG. 2 (SEQ ID NO:2),or is complementary to such encoding nucleic acid sequence, and remainsstably bound to it under at least moderate, and optionally, under highstringency conditions.

[0335] In another embodiment, the invention provides isolated PRO213polypeptide. In particular, the invention provides isolated nativesequence PRO213 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 295 of FIG. 2 (SEQ ID NO:2).

[0336] 2. PRO274

[0337] Applicants have identified a cDNA clone that encodes a novelpolypeptide, wherein the polypeptide is designated in the presentapplication as “PRO274”.

[0338] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO274 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO274polypeptide having amino acid residues 1 to 492 of FIG. 4 (SEQ ID NO:7),or is complementary to such encoding nucleic acid sequence, and remainsstably bound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA39987-1184 vector deposited on Apr. 21, 1998as ATCC 209786 which includes the nucleotide sequence encoding PRO274.

[0339] In another embodiment, the invention provides isolated PRO274polypeptide. In particular, the invention provides isolated nativesequence PRO274 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 492 of FIG. 4 (SEQ ID NO:7). Anadditional embodiment of the present invention is directed to anisolated extracellular domain of a PRO274 polypeptide. Optionally, thePRO274 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA39987-1184 vectordeposited on Apr. 21, 1998 as ATCC 209786.

[0340] In another embodiment, the invention provides three expressedsequence tags (EST) comprising the nucleotide sequences of SEQ ID NO:8(herein designated as DNA17873), SEQ ID NO:9 (herein designated asDNA36157) and SEQ ID NO:10 (herein designated as DNA28929) (see FIG.5-7, respectively).

[0341] 3. PRO300

[0342] Applicants have identified a cDNA clone that encodes a novelpolypeptide, wherein the polypeptide is designated in the presentapplication as “PRO300”.

[0343] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO300 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO300polypeptide having amino acid residues 1 to 457 of FIG. 9 (SEQ IDNO:19), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA40625-1189 vector deposited on Apr.21, 1998 as ATCC 209788 which includes the nucleotide sequence encodingPRO300.

[0344] In another embodiment, the invention provides isolated PRO300polypeptide. In particular, the invention provides isolated nativesequence PRO300 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 457 of FIG. 9 (SEQ ID NO:19). Anadditional embodiment of the present invention is directed to anisolated extracellular domain of a PRO300 polypeptide. Optionally, thePRO300 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA40625-1189 vectordeposited on Apr. 21, 1998 as ATCC 209788.

[0345] 4. PRO284

[0346] Applicants have identified a cDNA clone that encodes a noveltransmembrane polypeptide, wherein the polypeptide is designated in thepresent application as “PRO284”.

[0347] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO284 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO284polypeptide having amnno acid residues 1 to 285 of FIG. 11 (SEQ IDNO:28), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO284 polypeptide having amino acidresidues about 25 to 285 of FIG. 11 (SEQ ID NO:28) or 1 or about 25 to Xof FIG. 11 (SEQ ID NO:28), where X is any amino acid from 71 to 80 ofFIG. 11 (SEQ ID NO:28), or is complementary to such encoding nucleicacid sequence, and remains stably bound to it under at least moderate,and optionally, under high stringency conditions. The isolated nucleicacid sequence may comprise the cDNA insert of the DNA23318-1211 vectordeposited on Apr. 21, 1998 as ATCC 209787 which includes the nucleotidesequence encoding PRO284.

[0348] In another embodiment, the invention provides isolated PRO284polypeptide. In particular, the invention provides isolated nativesequence PRO284 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 285 of FIG. 11 (SEQ ID NO:28).Additional embodiments of the present invention are directed to isolatedPRO284 polypeptides comprising amino acids about 25 to 285 of FIG. 11(SEQ ID NO:28) or 1 or about 25 to X of FIG. 11 (SEQ ID NO:28), where Xis any amino acid from 71 to 80 of FIG. 11 (SEQ ID NO:28). Optionally,the PRO284 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA23318-1211 vectordeposited on Apr. 21, 1998 as ATCC 209787.

[0349] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA12982 which comprises thenucleotide sequence of SEQ ID NO:29.

[0350] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA15886 which comprises thenucleotide sequence of SEQ ID NO:30. 5. PRO296

[0351] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to the sarcoma-amplified protein SAS,wherein the polypeptide is designated in the present application as“PRO296”.

[0352] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO296 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO296polypeptide having amino acid residues 1 to 204 of FIG. 15 (SEQ IDNO:36), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO296 polypeptide having amino acidresidues about 35 to 204 of FIG. 15 (SEQ ID NO:36) or amino acid 1 orabout 35 to X of FIG. 15 (SEQ ID NO:36), where X is any amino acid from42 to 51 of FIG. 15 (SEQ ID NO:36), or is complementary to such encodingnucleic acid sequence, and remains stably bound to it under at leastmoderate, and optionally, under high stringency conditions. The isolatednucleic acid sequence may comprise the cDNA insert of the DNA39979-1213vector deposited on Apr. 21, 1998 as ATCC 209789 which includes thenucleotide sequence encoding PRO296.

[0353] In another embodiment, the invention provides isolated PRO296polypeptide. In particular, the invention provides isolated nativesequence PRO296 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 204 of FIG. 15 (SEQ ID NO:36).Additional embodiments of the present invention are directed to PRO296polypeptides comprising amino acids about 35 to 204 of FIG. 15 (SEQ IDNO:36) or amino acid 1 or about 35 to X of FIG. 15 (SEQ ID NO:36), whereX is any amino acid from 42 to 51 of FIG. 15 (SEQ ID NO:36). Optionally,the PRO296 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA39979-1213 vectordeposited on Apr. 21, 1998 as ATCC 209789.

[0354] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA23020 comprising thenucleotide sequence of SEQ ID NO:37.

[0355] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA21971 comprising thenucleotide sequence of SEQ ID NO:38.

[0356] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA29037 comprising thenucleotide sequence of SEQ ID NO:39. 6. PRO329

[0357] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to a high affinity immunoglobulin F_(c)receptor, wherein the polypeptide is designated in the presentapplication as “PRO329”.

[0358] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO329 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO329polypeptide having amino acid residues 1 to 359 of FIG. 20 (SEQ IDNO:45), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA40594-1233 vector deposited on Feb.5, 1998 as ATCC 209617 which includes the nucleotide sequence encodingPRO329.

[0359] In another embodiment, the invention provides isolated PRO329polypeptide. In particular, the invention provides isolated nativesequence PRO329 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 359 of FIG. 20 (SEQ ID NO:45).Optionally, the PRO329 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA40594-1233 vector deposited on Feb. 5, 1998 as ATCC 209617.

[0360] 7. PRO362

[0361] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to A33 antigen and HCAR membrane-boundprotein, wherein the polypeptide is designated in the presentapplication as “PRO362”.

[0362] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO362 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO362polypeptide having amino acid residues 1 to 321 of FIG. 22 (SEQ IDNO:52), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO362 polypeptide having aminoacid residues 1 to X of FIG. 22 (SEQ ID NO:52) where X is any amino acidfrom amino acid 271 to 280, or is complementary to such encoding nucleicacid sequence, and remains stably bound to it under at least moderate,and optionally, under high stringency conditions. The isolated nucleicacid sequence may comprise the cDNA insert of the DNA45416-1251 vectordeposited on Feb. 5, 1998 as ATCC 209620 which includes the nucleotidesequence encoding PRO362.

[0363] In another embodiment, the invention provides isolated PRO362polypeptide. In particular, the invention provides isolated nativesequence PRO362 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 321 of FIG. 22 (SEQ ID NO:52). Anadditional embodiment of the present invention is directed to anisolated extracellular domain of a PRO362 polypeptide comprising aminoacids 1 to X of the amino acid sequence shown in FIG. 22 (SEQ ID NO:52),wherein X is any amino acid from amino acid 271 to 280. Optionally, thePRO362 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA45416-1251 vectordeposited on Feb. 5, 1998 as ATCC 209620.

[0364] 8. PRO363

[0365] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to the cell surface receptor protein HCAR,wherein the polypeptide is designated in the present application as“PRO363”.

[0366] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO363 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO363polypeptide having amino acid residues 1 to 373 of FIG. 24 (SEQ IDNO:59), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding a PRO363 extracellular domainpolypeptide having amino acid residues 1 to X of FIG. 24 (SEQ ID NO:59)where X is any amino acid from amino acid 216 to amino acid 225, or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA45419-1252 vector deposited on Feb. 5, 1998 asATCC 209616 which includes the nucleotide sequence encoding PRO363.

[0367] In another embodiment, the invention provides isolated PRO363polypeptide. In particular, the invention provides isolated nativesequence PRO363 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 373 of FIG. 24 (SEQ ID NO:59). Anadditional embodiment of the present invention is directed to anisolated extracellular domain of a PRO363 polypeptide, wherein thatextracellular domain may comprise amino acids 1 to X of the sequenceshown in FIG. 24 (SEQ ID NO:59), where X is any amino acid from aminoacid 216 to 225. Optionally, the PRO363 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA45419-1252 vector deposited on Feb. 5, 1998 as ATCC 209616.

[0368] 9. PRO868

[0369] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to tumor necrosis factor receptor, whereinthe polypeptide is designated in the present application as “PRO868”.

[0370] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO868 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO868polypeptide having amino acid residues 1 to 655 of FIG. 26 (SEQ IDNO:64), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO868 polypeptide having aminoacid residues 1 to X of FIG. 26 (SEQ ID NO:64), where X is any aminoacid from amino acid 343 to 352 of the sequence shown in FIG. 26 (SEQ IDNO:64), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In yet another aspect, the isolatednucleic acid comprises DNA encoding the PRO868 polypeptide having aminoacid residues X to 655 of FIG. 26 (SEQ ID NO:64), where X is any aminoacid from amino acid 371 to 380 of the sequence shown in FIG. 26 (SEQ IDNO:64), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA52594-1270 vector deposited on Mar.17, 1998 as ATCC 209679 which includes the nucleotide sequence encodingPRO868.

[0371] In another embodiment, the invention provides isolated PRO868polypeptide. In particular, the invention provides isolated nativesequence PRO868 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 655 of FIG. 26 (SEQ ID NO:64). Inanother aspect, the isolated PRO868 polypeptide comprises amino acidresidues 1 to X of FIG. 26 (SEQ ID NO:64), where X is any amino acidfrom amino acid 343 to 352 of the sequence shown in FIG. 26 (SEQ IDNO:64). In yet another aspect, the PRO868 polypeptide comprises aminoacid residues X to 655 of FIG. 26 (SEQ ID NO:64), where X is any aminoacid from amino acid 371 to 380 of the sequence shown in FIG. 26 (SEQ IDNO:64). Optionally, the PRO868 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA52594-1270 vector deposited on Mar. 17, 1998 as ATCC 209679.

[0372] 10. PRO382

[0373] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to serine proteases, wherein the polypeptideis designated in the present application as “PRO382”.

[0374] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO382 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO382polypeptide having amino acid residues 1 to 453 of FIG. 28 (SEQ IDNO:69), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA45234-1277 vector deposited on Mar.5, 1998 as ATCC 209654 which includes the nucleotide sequence encodingPRO382.

[0375] In another embodiment, the invention provides isolated PRO382polypeptide. In particular, the invention provides isolated nativesequence PRO382 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 453 of FIG. 28 (SEQ ID NO:69). Anadditional embodiment of the present invention is directed to anisolated extracellular domain of a PRO382 polypeptide, with or withoutthe signal peptide. Optionally, the PRO382 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA45234-1277 vector deposited on Mar. 5, 1998 as ATCC 209654.

[0376] 11. PRO545

[0377] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to meltrin, wherein the polypeptide isdesignated in the present application as “PRO545”.

[0378] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO545 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO545polypeptide having amino acid residues 1 to 735 of FIG. 30 (SEQ IDNO:74), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Mar. 5, 1998 as ATCC209655 which includes the nucleotide sequence encoding PRO545.

[0379] In another embodiment, the invention provides isolated PRO545polypeptide. In particular, the invention provides isolated nativesequence PRO545 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 735 of FIG. 30 (SEQ ID NO:74). Anadditional embodiment of the present invention is directed to anisolated extracellular domain of a PRO545 polypeptide. Optionally, thePRO545 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Mar.5, 1998 as ATCC 209655.

[0380] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA13217 comprising thenucleotide sequence of SEQ ID NO:75 (FIG. 31).

[0381] 12. PRO617

[0382] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to CD24, wherein the polypeptide isdesignated in the present application as “PRO617”.

[0383] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO617 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO617polypeptide having amino acid residues 1 to 67 of FIG. 33 (SEQ IDNO:85), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA48309-1280 vector deposited on Mar.5, 1998 as ATCC 209656 which includes the nucleotide sequence encodingPRO617.

[0384] In another embodiment, the invention provides isolated PRO617polypeptide. In particular, the invention provides isolated nativesequence PRO617 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 67 of FIG. 33 (SEQ ID NO:85).Optionally, the PRO617 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA48309-1280 vector deposited on Mar. 5, 1998 as ATCC 209656.

[0385] 13. PRO700

[0386] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to protein disulfide isomerase,wherein the polypeptide is designated in the present application as“PRO700”.

[0387] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO700 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO700polypeptide having amino acid residues 1 to 432 of FIG. 35 (SEQ IDNO:90), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO700 polypeptide having aminoacid residues from about 34 to 432 of FIG. 35 (SEQ ID NO:90), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the vector deposited on Mar. 31, 1998 as ATCC 209721which includes the nucleotide sequence encoding PRO700.

[0388] In another embodiment, the invention provides isolated PRO700polypeptide. In particular, the invention provides isolated nativesequence PRO700 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 432 of FIG. 35 (SEQ ID NO:90). Inanother embodiment, the invention provides an isolated PRO700polypeptide absent the signal sequence, which includes an amino acidsequence comprising residues from about 34 to 432 of FIG. 35 (SEQ IDNO:90). Optionally, the PRO700 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of the vectordeposited on Mar. 31, 1998 as ATCC 209721.

[0389] 14. PRO702

[0390] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to conglutinin, wherein the polypeptide isdesignated in the present application as “PRO702”.

[0391] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO702 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO702polypeptide having amino acid residues 1 to 277 of FIG. 37 (SEQ IDNO:97), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO702 polypeptide having aminoacid residues 26 to 277 of FIG. 37 (SEQ ID NO:97), or is complementaryto such encoding nucleic acid sequence, and remains stably bound to itunder at least moderate, and optionally, under high stringencyconditions. The isolated nucleic acid sequence may comprise the cDNAinsert of the DNA50980-1286 vector deposited on Mar. 31, 1998 as ATCC209717 which includes the nucleotide sequence encoding PRO702.

[0392] In another embodiment, the invention provides isolated PRO702polypeptide. In particular, the invention provides isolated nativesequence PRO702 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 277 of FIG. 37 (SEQ ID NO:97). Anadditional embodiment of the present invention is directed to anisolated PRO702 polypeptide comprising amino acid residues 26 to 277 ofFIG. 37 (SEQ ID NO:97). Optionally, the PRO702 polypeptide is obtainedor is obtainable by expressing the polypeptide encoded by the cDNAinsert of the DNA50980-1286 vector deposited on Mar. 31, 1998 as ATCC209717.

[0393] 15. PRO703

[0394] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to VLCAS, wherein the polypeptideis designated in the present application as “PRO703”.

[0395] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO703 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO703polypeptide having amino acid residues 1 to 730 of FIG. 39 (SEQ IDNO:102), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO703 polypeptide having aminoacid residues from about 43 to 730 of FIG. 39 (SEQ ID NO:102), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA50913-1287 vector deposited on Mar. 31, 1998as ATCC 209716 which includes the nucleotide sequence encoding PRO703.

[0396] In another embodiment, the invention provides isolated PRO703polypeptide. In particular, the invention provides isolated nativesequence PRO703 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 730 of FIG. 39 (SEQ ID NO:102).In another embodiment, the invention provides an isolated PRO703polypeptide absent the signal sequence, which includes an amino acidsequence comprising residues from about 43 to 730 of FIG. 30 (SEQ IDNO:102). Optionally, the PRO730 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA50913-1287 vector deposited on Mar. 31, 1998 as ATCC 209716.

[0397] 16. PRO705

[0398] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to K-glypican, wherein the polypeptide isdesignated in the present application as “PRO705”.

[0399] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO705 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO705polypeptide having amino acid residues 1 to 555 of FIG. 41 (SEQ IDNO:109), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO705 polypeptide having aminoacid residues about 24 to 555 of FIG. 41 (SEQ ID NO:109), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA50914-1289 vector deposited on Mar. 31, 1998as ATCC 209722 which includes the nucleotide sequence encoding PRO705.

[0400] In another embodiment, the invention provides isolated PRO705polypeptide. In particular, the invention provides isolated nativesequence PRO705 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 555 of FIG. 41 (SEQ ID NO:109).An additional embodiment of the present invention is directed to anisolated PRO705 polypeptide comprising amino acid residues about 24 to555 of FIG. 41 (SEQ ID NO:109). Optionally, the PRO705 polypeptide isobtained or is obtainable by expressing the polypeptide encoded by thecDNA insert of the DNA50914-1289 vector deposited on Mar. 31, 1998 asATCC 209722.

[0401] 17. PRO708

[0402] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to the aryl sulfatases, wherein thepolypeptide is designated in the present application as “PRO708”.

[0403] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO708 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO708polypeptide having amino acid residues 1 to 515 of FIG. 43 (SEQ IDNO:114), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA48296-1292 vector deposited on Mar.11, 1998 as ATCC 209668 which includes the nucleotide sequence encodingPRO708.

[0404] In another embodiment, the invention provides isolated PRO708polypeptide. In particular, the invention provides isolated nativesequence PRO708 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 515 of FIG. 43 (SEQ ID NO:114).Another embodiment is directed to a PRO708 polypeptide comprisingresidues 38-515 of the amino acid sequence shown in FIG. 43 (SEQ IDNO:114). Optionally, the PRO708 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA48296-1292 vector deposited on Mar. 11, 1998 as ATCC 209668.

[0405] 18. PRO320

[0406] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to fibulin, wherein the polypeptide isdesignated in the present application as “PRO320”.

[0407] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO320 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO320polypeptide having amino acid residues 1 to 338 of FIG. 45 (SEQ IDNO:119), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Mar. 11, 1998 asATCC 209670 which includes the nucleotide sequence encoding PRO320.

[0408] In another embodiment, the invention provides isolated PRO320polypeptide. In particular, the invention provides isolated nativesequence PRO320 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 338 of FIG. 45 (SEQ ID NO:119).Optionally, the PRO320 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of the vectordeposited on Mar. 11, 1998 as ATCC 209670.

[0409] 19. PRO324

[0410] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to oxidoreductases, wherein the polypeptideis designated in the present application as “PRO324”.

[0411] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO324 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO324polypeptide having amino acid residues 1 to 289 of FIG. 47 (SEQ IDNO:124), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO324 polypeptide having amino acidresidues 1 or about 32 to X of FIG. 47 (SEQ ID NO:124), where X is anyamino acid from 131 to 140, or is complementary to such encoding nucleicacid sequence, and remains stably bound to it under at least moderate,and optionally, under high stringency conditions. The isolated nucleicacid sequence may comprise the cDNA insert of the DNA36343-1310 vectordeposited on Mar. 30, 1998 as ATCC 209718 which includes the nucleotidesequence encoding PRO324.

[0412] In another embodiment, the invention provides isolated PRO324polypeptide. In particular, the invention provides isolated nativesequence PRO324 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 289 of FIG. 47 (SEQ ID NO:124).The invention also provides isolated PRO324 polypeptide comprisingresidues 1 or about 32 to X of FIG. 47 (SEQ ID NO:124), wherein X is anyamino acid from about 131-140. Optionally, the PRO324 polypeptide isobtained or is obtainable by expressing the polypeptide encoded by thecDNA insert of the DNA36343-1310 vector deposited on Mar. 30, 1998 asATCC 209718.

[0413] 20. PRO351

[0414] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to prostasin, wherein thepolypeptide is designated in the present application as “PRO351”.

[0415] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO351 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO351polypeptide having amino acid residues 1 to 571 of FIG. 49 (SEQ IDNO:132), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO351 polypeptide having aminoacid residues about 16 to 571 of FIG. 49 (SEQ ID NO:132), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA40571-1315 vector deposited on Apr. 21, 1998as ATCC 209784 which includes the nucleotide sequence encoding PRO351.

[0416] In another embodiment, the invention provides isolated PRO351polypeptide. In particular, the invention provides isolated nativesequence PRO351 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 571 of FIG. 49 (SEQ ID NO:132).In another embodiment, the invention provides an isolated PRO351polypeptide absent the signal sequence, which includes an amino acidsequence comprising residues from about 16 to 571 of FIG. 49 (SEQ IDNO:132). Optionally, the PRO351 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA40571-1315 vector deposited on Apr. 21, 1998 as ATCC 209784.

[0417] 21. PRO352

[0418] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to butyrophilin, wherein the polypeptide isdesignated in the present application as “PRO352”.

[0419] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO352 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO352polypeptide having amino acid residues 1 to 316 of FIG. 51 (SEQ IDNO:137), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO352 polypeptide having amino acidresidues of about 29 to 316 of FIG. 51 (SEQ ID NO:137), or 1 or about 29to X of FIG. 51, where X is any amino acid from 246 to 255, or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA41386-1316 vector deposited on Mar. 26, 1998as ATCC 209703 which includes the nucleotide sequence encoding PRO352.

[0420] In another embodiment, the invention provides isolated PRO352polypeptide. In particular, the invention provides isolated nativesequence PRO352 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 316 of FIG. 51 (SEQ ID NO:137).In other embodiments, the invention provides isolated PRO352 polypeptidecomprising residues about 29 to 316 of FIG. 51 (SEQ ID NO:137) and 1 orabout 29 to X of FIG. 51 (SEQ ID NO:137), wherein X is any amino acidfrom 246 to 255. Optionally, the PRO352 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA41386-1316 vector deposited on Mar. 26, 1998 as ATCC 209703.

[0421] 22. PRO381

[0422] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to immunophilin proteins, wherein thepolypeptide is designated in the present application as “PRO381”.

[0423] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO381 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO381polypeptide having amino acid residues 1 to 211 of FIG. 53 (SEQ IDNO:145), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO381 polypeptide having aminoacid residues about 21 to 211 of FIG. 53 (SEQ ID NO:145), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA44194-1317 vector deposited on Apr. 28, 1998as ATCC 209808 which includes the nucleotide sequence encoding PRO381.

[0424] In another embodiment, the invention provides isolated PRO381polypeptide. In particular, the invention provides isolated nativesequence PRO381 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 211 of FIG. 53 (SEQ ID NO:145).Another embodiment is directed to a PRO381 polypeptide comprising aminoacids about 21 to 211 of FIG. 53 (SEQ ID NO:145). Optionally, the PRO381polypeptide is obtained or is obtainable by expressing the polypeptideencoded by the cDNA insert of the DNA44194-1317 vector deposited on Apr.28, 1998 as ATCC 209808.

[0425] 23. PRO386

[0426] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to the beta-2 subunit of a sodium channel,wherein the polypeptide is designated in the present application as“PRO386”.

[0427] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO386 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO386polypeptide having amino acid residues 1 to 215 of FIG. 55 (SEQ IDNO:150), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO386 polypeptide having aminoacid residues about 21 to 215 of FIG. 55 (SEQ ID NO:150) or 1 or about21 to X, where X is any amino acid from 156 to 165 of FIG. 55 (SEQ IDNO:150), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA45415-1318 vector deposited on Apr.28, 1998 as ATCC 209810 which includes the nucleotide sequence encodingPRO386.

[0428] In another embodiment, the invention provides isolated PRO386polypeptide. In particular, the invention provides isolated nativesequence PRO386 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 215 of FIG. 55 (SEQ ID NO:150).Other embodiments of the present invention are directed to PRO386polypeptides comprising amino acids about 21 to 215 of FIG. 55 (SEQ IDNO:150) and 1 or about 21 to X of FIG. 55 (SEQ ID NO:150), wherein X isany amino acid from 156 to 165 of FIG. 55 (SEQ ID NO:150). Optionally,the PRO386 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA45415-1318 vectordeposited on Apr. 28, 1998 as ATCC 209810.

[0429] In another embodiment, the invention provides an expressedsequence tag (EST) comprising the nucleotide sequence of SEQ ID NO:151which corresponds to an EST designated herein as DNA23350.

[0430] In another embodiment, the invention provides an expressedsequence tag (EST) comprising the nucleotide sequence of SEQ ID NO:152which corrsponds to an EST designated herein as DNA23536.

[0431] 24. PRO540

[0432] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to LCAT, wherein the polypeptideis designated in the present application as “PRO540”.

[0433] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO540 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO540polypeptide having amino acid residues 1 to 412 of FIG. 59 (SEQ IDNO:157), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO540 polypeptide having aminoacid residues about 29 to 412 of FIG. 59 (SEQ ID NO:157), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA44189-1322 vector deposited on Mar. 26, 1998as ATCC 209699 which includes the nucleotide sequence encoding PRO540.

[0434] In another embodiment, the invention provides isolated PRO540polypeptide. In particular, the invention provides isolated nativesequence PRO540 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 412 of FIG. 59 (SEQ ID NO:157).The invention also provides isolated PRO540 polypeptide, which in oneembodiment, includes an amino acid sequence comprising residues about 29to 412 of FIG. 59 (SEQ ID NO:157). Optionally, the PRO540 polypeptide isobtained or is obtainable by expressing the polypeptide encoded by thecDNA insert of the DNA44189-1322 vector deposited on Mar. 26, 1998 asATCC 209699.

[0435] 25. PRO615

[0436] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to synaptogyrin, wherein thepolypeptide is designated in the present application as “PRO615”.

[0437] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO615 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO615polypeptide having amino acid residues 1 to 224 of FIG. 61 (SEQ IDNO:162), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO615 polypeptide having aminoacid residues X to 224 of FIG. 61 (SEQ ID NO:162), where X is any aminoacid from 157 to 166, or is complementary to such encoding nucleic acidsequence, and remains stably bound to it under at least moderate, andoptionally, under high stringency conditions. The isolated nucleic acidsequence may comprise the cDNA insert of the DNA48304-1323 vectordeposited on Apr. 28, 1998 as ATCC 209811 which includes the nucleotidesequence encoding PRO615.

[0438] In another embodiment, the invention provides isolated PRO615polypeptide. In particular, the invention provides isolated nativesequence PRO615 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 224 of FIG. 61 (SEQ ID NO:162).An additional embodiment of the present invention is directed to anisolated extracellular domain of a PRO615 polypeptide which comprisesamino acid residues X to 224 of FIG. 61 (SEQ ID NO:162), where X is anyamino acid from 157 to 166 of FIG. 61 (SEQ ID NO:162). Optionally, thePRO615 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA48304-1323 vectordeposited on Apr. 28, 1998 as ATCC 209811.

[0439] 26. PRO618

[0440] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to enteropeptidase, wherein thepolypeptide is designated in the present application as “PRO618”.

[0441] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO618 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO618polypeptide having amino acid residues 1 to 802 of FIG. 63 (SEQ IDNO:169), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding an isolated extracellular domain ofa PRO618 polypeptide having amino acid residues X to 802 of FIG. 63 (SEQID NO:169), where X is any amino acid from 63 to 72 of FIG. 63 (SEQ IDNO:169), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the DNA49152-1324 vector deposited on Apr.28, 1998 as ATCC 209813 which includes the nucleotide sequence encodingPRO618.

[0442] In another embodiment, the invention provides isolated PRO618polypeptide. In particular, the invention provides isolated nativesequence PRO618 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 802 of FIG. 63 (SEQ ID NO:169).An additional embodiment of the present invention is directed to anisolated extracellular domain of a PRO618 polypeptide comprising aminoacid X to 802 where X is any amino acid from 63 to 72 of FIG. 63 (SEQ IDNO:169). Optionally, the PRO618 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA49152-1324 vector deposited on Apr. 28, 1998 as ATCC 209813.

[0443] In another embodiment, the invention provides an expressedsequence tag (ESf) comprising the nucleotide sequence of SEQ ID NO:170,designated herein as DNA35597 (see FIG. 64).

[0444] 27. PRO719

[0445] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to lipoprotein lipase H, wherein thepolypeptide is designated in the present application as “PRO719”.

[0446] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO719 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO719polypeptide having amino acid residues 1 to 354 of FIG. 66 (SEQ IDNO:178), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO719 polypeptide having aminoacid residues about 17 to 354 of FIG. 66 (SEQ ID NO:178), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA49646-1327 vector deposited on Mar. 26, 1998as ATCC 209705 which includes the nucleotide sequence encoding PRO719.

[0447] In another embodiment, the invention provides isolated PRO719polypeptide. In particular, the invention provides isolated nativesequence PRO719 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 354 of FIG. 66 (SEQ ID NO:178).In another embodiment, the invention provides isolated PRO719polypeptide which comprises residues about 17 to 354 of FIG. 66 (SEQ IDNO:178). Optionally, the PRO719 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA49646-1327 vector deposited on Mar. 26, 1998 as ATCC 209705.

[0448] 28. PRO724

[0449] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to the LDL receptor, wherein the polypeptideis designated in the present application as “PRO724”.

[0450] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO724 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO724polypeptide having amino acid residues 1 to 713 of FIG. 68 (SEQ IDNO:183), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding a soluble PRO724 polypeptide havingamino acid residues 1 to X of FIG. 68 (SEQ ID NO:183) where X is anyamino acid from amino acid 437 to 446, or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theabove two polypeptides may either possess or not possess the signalpeptide. The isolated nucleic acid sequence may comprise the cDNA insertof the DNA49631-1328 vector deposited on Apr. 28, 1998 as ATCC 209806which includes the nucleotide sequence encoding PRO724.

[0451] In another embodiment, the invention provides isolated PRO724polypeptide. In particular, the invention provides isolated nativesequence PRO724 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 713 of FIG. 68 (SEQ ID NO:183).In another embodiment, the invention provides isolated soluble PRO724polypeptide. In particular, the invention provides isolated solublePRO724 polypeptide, which in one embodiment, includes an amino acidsequence comprising residues 1 to X of FIG. 68 (SEQ ID NO:183), where Xis any amino acid from 437 to 446 of the sequence shown in FIG. 68 (SEQID NO:183). Optionally, the PRO724 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA49631-1328 vector deposited on Apr. 28, 1998 as ATCC 209806.

[0452] 29. PRO772

[0453] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to A4 protein, wherein the polypeptide isdesignated in the present application as “PRO772”.

[0454] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO772 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO772polypeptide having amino acid residues 1 to 152 of FIG. 70 (SEQ IDNO:190), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO772 polypeptide having amino acidresidues 1 to X of FIG. 70 (SEQ ID NO:190), where X is any amino acidfrom 21 to 30 of FIG. 70 (SEQ ID NO:190), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA49645-1347 vector deposited on Apr. 28, 1998 as ATCC 209809 whichincludes the nucleotide sequence encoding PRO772.

[0455] In another embodiment, the invention provides isolated PRO772polypeptide. In particular, the invention provides isolated nativesequence PRO772 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 152 of FIG. 70 (SEQ ID NO:190).Additional embodiments of the present invention are directed to PRO772polypeptides comprising amino acids 1 to X of FIG. 70 (SEQ ID NO:190),where X is any amino acid from 21 to 30 of FIG. 70 (SEQ ID NO:190).Optionally, the PRO772 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA49645-1347 vector deposited on Apr. 28, 1998 as ATCC 209809.

[0456] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA43509 comprising thenucleotide sequence of SEQ ID NO:191 (FIG. 71).

[0457] 30. PRO852

[0458] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to various protease enzymes, wherein thepolypeptide is designated in the present application as “PRO852”.

[0459] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO852 polypeptide. hl oneaspect, the isolated nucleic acid comprises DNA encoding the PRO852polypeptide having amino acid residues 1 to 518 of FIG. 73 (SEQ IDNO:196), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO852 polypeptide having amino acidresidues about 21 to 518 of FIG. 73 (SEQ ID NO:196) or 1 or about 21 toX of FIG. 73 (SEQ ID NO:196) where X is any amino acid from amino acid461 to amino acid 470 of FIG. 73 (SEQ ID NO:196), or is complementary tosuch encoding nucleic acid sequence, and remains stably bound to itunder at least moderate, and optionally, under high stringencyconditions. The isolated nucleic acid sequence may comprise the cDNAinsert of the DNA45493-1349 vector deposited on Apr. 28, 1998 as ATCC209805 which includes the nucleotide sequence encoding PRO852.

[0460] In another embodiment, the invention provides isolated PRO852polypeptide. In particular, the invention provides isolated nativesequence PRO852 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 518 of FIG. 73 (SEQ ID NO:196).In other embodiments, the PRO852 comprises amino acids about 21 to aminoacid 518 of FIG. 73 (SEQ ID NO:196) or amino acids 1 or about 21 to X ofFIG. 73 (SEQ ID NO:196), where X is any amino acid from amino acid 461to amino acid 470 of FIG. 73 (SEQ ID NO:196). Optionally, the PRO852polypeptide is obtained or is obtainable by expressing the polypeptideencoded by the cDNA insert of the DNA45493-1349 vector deposited on Apr.28, 1998 as ATCC 209805.

[0461] 31. PRO853

[0462] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to reductase, wherein thepolypeptide is designated in the present application as “PRO853”.

[0463] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO853 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO853polypeptide having amino acid residues 1 to 377 of FIG. 75 (SEQ IDNO:206), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO853 polypeptide having aminoacid residues about 17 to 377 of FIG. 75 (SEQ ID NO:206), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA48227-1350 vector deposited on Apr. 28, 1998as ATCC 209812 which includes the nucleotide sequence encoding PRO853.

[0464] In another embodiment, the invention provides isolated PRO853polypeptide. In particular, the invention provides isolated nativesequence PRO853 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 377 of FIG. 75 (SEQ ID NO:206).In another embodiment, the invention provides an isolated PRO853polypeptide absent the signal sequence, which includes an amino acidsequence comprising residues from about 17 to 377 of FIG. 75 (SEQ IDNO:206). Optionally, the PRO853 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA48227-1350 vector deposited on Apr. 28, 1998 as ATCC 209812.

[0465] 32. PRO860

[0466] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to neurofascin, wherein thepolypeptide is designated in the present application as “PRO860”.

[0467] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO860 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO860polypeptide having amino acid residues 1 to 985 of FIG. 77 (SEQ IDNO:211), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO860 polypeptide having aminoacid residues 1 to X of FIG. 77 (SEQ ID NO:211), where X is any aminoacid from 443-452 of FIG. 77 (SEQ ID NO:211), or is complementary tosuch encoding nucleic acid sequence, and remains stably bound to itunder at least moderate, and optionally, under high stringencyconditions. The isolated nucleic acid sequence may comprise the cDNAinsert of the DNA41404-1352 vector deposited on May 6, 1998 as ATCC209844 which includes the nucleotide sequence encoding PRO860.

[0468] In another embodiment, the invention provides isolated PRO860polypeptide. In particular, the invention provides isolated nativesequence PRO860 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 985 of FIG. 77 (SEQ ID NO:211).In another embodiment, the invention provides an isolated PRO860polypeptide which includes an amino acid sequence comprising residues 1to X of FIG. 77 (SEQ ID NO:211), where X is any amino acid residue from443 to 452 of FIG. 77 (SEQ ID NO:211). Optionally, the PRO860polypeptide is obtained or is obtainable by expressing the polypeptideencoded by the cDNA insert of the DNA41404-1352 vector deposited on May6, 1998 as ATCC 209844.

[0469] 33. PRO846

[0470] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to CMRF35, wherein thepolypeptide is designated in the present application as “PRO846”.

[0471] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO846 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO846polypeptide having amino acid residues 1 to 332 of FIG. 79 (SEQ IDNO:216), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO846 polypeptide having amino acidresidues about 18 to 332 of FIG. 79 (SEQ ID NO:216) or 1 or about 18 toX of SEQ ID NO:216, where X is any amino acid from 243 to 252 of FIG. 79(SEQ ID NO:216), or is complementary to such encoding nucleic acidsequence, and remains stably bound to it under at least moderate, andoptionally, under high stringency conditions. The isolated nucleic acidsequence may comprise the cDNA insert of the DNA44196-1353 vectordeposited on May 6, 1998 as ATCC 209847 which includes the nucleotidesequence encoding PRO846.

[0472] In another embodiment, the invention provides isolated PRO846polypeptide. In particular, the invention provides isolated nativesequence PRO846 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 332 of FIG. 79 (SEQ ID NO:216).In other embodiments, the invention provides an isolated PRO846polypeptide absent the signal sequence, which includes an amino acidsequence comprising residues from about 18 to 332 of FIG. 79 (SEQ IDNO:216). Additional embodiments of the present invention are directed toan isolated PRO846 polypeptide comprising amino acid 1 or about 18 to Xof FIG. 79 (SEQ ID NO:216), where X is any amino acid from 243 to 252 ofFIG. 79 (SEQ ID NO:216). Optionally, the PRO846 polypeptide is obtainedor is obtainable by expressing the polypeptide encoded by the cDNAinsert of the DNA44196-1353 vector deposited on May 6, 1998 as ATCC209847.

[0473] 34. PRO862

[0474] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to lysozyme, wherein thepolypeptide is designated in the present application as “PRO862”.

[0475] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO862 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO862polypeptide having amino acid residues 1 to 146 of FIG. 81 (SEQ IDNO:221), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO862 polypeptide having aminoacid residues about 19 to 146 of FIG. 81 (SEQ ID NO:221), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA52187-1354 vector deposited on May 6, 1998 asATCC 209845 which includes the nucleotide sequence encoding PRO862.

[0476] In another embodiment, the invention provides isolated PRO862polypeptide. In particular, the invention provides isolated nativesequence PRO862 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 146 of FIG. 81 (SEQ ID NO:221).in another embodiment, the invention provides an isolated PRO862polypeptide absent the signal sequence, which includes an amino acidsequence comprising residues from about 19 to 146 of FIG. 81 (SEQ IDNO:221). Optionally, the PRO862 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA52187-1354 vector deposited on May 6, 1998 as ATCC 209845.

[0477] 35. PRO864

[0478] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence similarity to Wnt-4, wherein the polypeptideis designated in the present application as “PRO864”.

[0479] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO864 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO864polypeptide having amino acid residues 1 to 351 of FIG. 83 (SEQ IDNO:226), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO864 polypeptide having aminoacid residues about 23 to 351 of FIG. 83 (SEQ ID NO:226), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA48328-1355 vector deposited on May 6, 1998 asATCC 209843 which includes the nucleotide sequence encoding PRO864.

[0480] In another embodiment, the invention provides isolated PRO864polypeptide. In particular, the invention provides isolated nativesequence PRO864 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 351 of FIG. 83 (SEQ ID NO:226).In another embodiment, the invention provides an isolated PRO864polypeptide absent the signal sequence, which includes an amino acidsequence comprising residues from about 23 to 351 of FIG. 83 (SEQ IDNO:226). Optionally, the PRO864 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA48328-1355 vector deposited on May 6, 1998 as ATCC 209843 .

[0481] 36. PRO792

[0482] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to CD23, wherein the polypeptide isdesignated in the present application as “PRO792”.

[0483] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO792 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO792polypeptide having amino acid residues 1 to 293 of FIG. 85 (SEQ IDNO:231), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO792 polypeptide having aminoacid residues X to 293 of FIG. 85 (SEQ ID NO:231) where X is any aminoacid from 50 to 59 of FIG. 85 (SEQ ID NO:231), or is complementary tosuch encoding nucleic acid sequence, and remains stably bound to itunder at least moderate, and optionally, under high stringencyconditions. The isolated nucleic acid sequence may comprise the cDNAinsert of the DNA56352-1358 vector deposited on May 6, 1998 as ATCC209846 which includes the nucleotide sequence encoding PRO792.

[0484] In another embodiment, the invention provides isolated PRO792polypeptide. In particular, the invention provides isolated nativesequence PRO792 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 293 of FIG. 85 (SEQ ID NO:231).An additional embodiment of the present invention is directed to PRO792polypeptide comprising amino acids X to 293 of FIG. 85 (SEQ ID NO:231),where X is any amino acid from 50 to 59 of FIG. 85 (SEQ ID NO:231).Optionally, the PRO792 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA56352-1358 vector deposited on May 6, 1998 as ATCC 209846.

[0485] 37. PRO866

[0486] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to mindin and spondin proteins, wherein thepolypeptide is designated in the present application as “PRO866”.

[0487] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO866 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO866polypeptide having amino acid residues 1 to 331 of FIG. 87 (SEQ IDNO:236), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another aspect, the isolatednucleic acid comprises DNA encoding the PRO866 polypeptide having aminoacid residues about 27 to 229 of FIG. 87 (SEQ ID NO:236), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA53971-1359 vector deposited on Apr. 7, 1998 asATCC 209750 which includes the nucleotide sequence encoding PRO866.

[0488] In another embodiment, the invention provides isolated PRO866polypeptide. In particular, the invention provides isolated nativesequence PRO866 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 331 of FIG. 87 (SEQ ID NO:236).Another embodiment of the present invention is directed to PRO866polypeptides comprising amino acids about 27 to 331 of FIG. 87 (SEQ IDNO:236). Optionally, the PRO866 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA53971-1359 vector deposited on Apr. 7, 1998 as ATCC 209750.

[0489] 38. PRO871

[0490] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to CyP-60, wherein the polypeptide isdesignated in the present application as “PRO871”.

[0491] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO871 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO871polypeptide having amino acid residues 1 to 472 of FIG. 89 (SEQ IDNO:245), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO871 polypeptide having amino acidresidues about 22 to 472 of FIG. 89 (SEQ ID NO:245), or is complementaryto such encoding nucleic acid sequence, and remains stably bound to itunder at least moderate, and optionally, under high stringencyconditions. The isolated nucleic acid sequence may comprise the cDNAinsert of the DNA50919-1361 vector deposited on May 6, 1998 as ATCC209848 which includes the nucleotide sequence encoding PRO871.

[0492] In another embodiment, the invention provides isolated PRO871polypeptide. In particular, the invention provides isolated nativesequence PRO871 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 472 of FIG. 89 (SEQ ID NO:245).An additional embodiment of the present invention is directed to PRO871polypeptides comprising amino acids about 22 to 472 of FIG. 89 (SEQ IDNO:245). Optionally, the PRO871 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA50919-1361 vector deposited on May 6, 1998 as ATCC 209848.

[0493] 39. PRO873

[0494] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to carboxylesterase, wherein the polypeptideis designated in the present application as “PRO873”.

[0495] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO873 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO873polypeptide having amino acid residues 1 to 545 of FIG. 91 (SEQ IDNO:254), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO873 polypeptide having amino acidresidues about 30 to about 545 of FIG. 91 (SEQ ID NO:254), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA44179-1362 vector deposited on May 6, 1998 asATCC 209851 which includes the nucleotide sequence encoding PRO873.

[0496] In another embodiment, the invention provides isolated PRO873polypeptide. In particular, the invention provides isolated nativesequence PRO873 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 545 of FIG. 91 (SEQ ID NO:254).Additional embodiments of the present invention are directed to PRO873polypeptides comprising amino acids about 30 to about 545 of FIG. 91(SEQ ID NO:254). Optionally, the PRO873 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA44179-1362 vector deposited on May 6, 1998 as ATCC 209851.

[0497] 40. PRO940

[0498] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to CD33 and OB binding protein-2, whereinthe polypeptide is designated in the present application as “PRO940”.

[0499] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO940 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO940polypeptide having amino acid residues 1 to 544 of FIG. 93 (SEQ IDNO:259), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO940 polypeptide having amino acidresidues about 16 to 544 of FIG. 93 (SEQ ID NO:259) or 1 or about 16 toX of FIG. 93 (SEQ ID NO:259), where X is any amino acid from 394 to 403of FIG. 93 (SEQ ID NO:259), or is complementary to such encoding nucleicacid sequence, and remains stably bound to it under at least moderate,and optionally, under high stringency conditions. The isolated nucleicacid sequence may comprise the cDNA insert of the DNA54002-1367 vectordeposited on Apr. 7, 1998 as ATCC 209754 which includes the nucleotidesequence encoding PRO940.

[0500] In another embodiment, the invention provides isolated PRO940polypeptide. In particular, the invention provides isolated nativesequence PRO940 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 544 of FIG. 93 (SEQ ID NO:259).Other embodiments of the present invention are directed to PRO940polypeptides comprising amino acids about 16 to 544 of FIG. 93 (SEQ IDNO:259) or 1 or about 16 to X of FIG. 93 (SEQ ID NO:259), where X is anyamino acid from 394 to 403 of FIG. 93 (SEQ ID NO:259). Optionally, thePRO940 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA54002-1367 vectordeposited on Apr. 7, 1998 as ATCC 209754.

[0501] 41. PRO941

[0502] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to a cadherin protein, wherein thepolypeptide is designated in the present application as “PRO941”.

[0503] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO941 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO941polypeptide having amino acid residues 1 to 772 of FIG. 95 (SEQ IDNO:264), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO941 polypeptide having amino acidresidues about 22 to 772 of FIG. 95 (SEQ ID NO:264) or 1 or about 22 toX of FIG. 95 (SEQ ID NO:264), where X is any amino acid from 592 to 601of FIG. 95 (SEQ ID NO:264), or is complementary to such encoding nucleicacid sequence, and remains stably bound to it under at least moderate,and optionally, under high stringency conditions. The isolated nucleicacid sequence may comprise the cDNA insert of the DNA53906-1368 vectordeposited on Apr. 7, 1998 as ATCC 209747 which includes the nucleotidesequence encoding PRO941.

[0504] In another embodiment, the invention provides isolated PRO941polypeptide. In particular, the invention provides isolated nativesequence PRO941 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 772 of FIG. 95 (SEQ ID NO:264).Additional embodiments of the present invention are directed to PRO941polypeptides which comprise amino acid about 21 to 772 of FIG. 95 (SEQID NO:264) or 1 or about 22 to X of FIG. 95 (SEQ ID NO:264), where X isany amino acid from 592 to 601 of FIG. 95 (SEQ ID NO:264). Optionally,the PRO941 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the DNA53906-1368 vectordeposited on Apr. 7, 1998 as ATCC 209747.

[0505] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA6415 comprising thenucleotide sequence of FIG. 96 (SEQ ID NO:265).

[0506] 42. PRO944

[0507] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to Clostridium perfringens enterotoxinreceptor (CPE-R), wherein the polypeptide is designated in the presentapplication as “PRO944”.

[0508] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO944 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO944polypeptide having amino acid residues 1 to 211 of FIG. 98 (SEQ IDNO:270), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO944 polypeptide having amino acidresidues about 22 to 229 of FIG. 98 (SEQ ID NO:270) or amino acid 1 orabout 22 to X of FIG. 98 (SEQ ID NO:270) where X is any amino acid from77 to 80 of FIG. 98 (SEQ ID NO:270), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA52185-1370 vector deposited on May 14, 1998 as ATCC 209861 whichincludes the nucleotide sequence encoding PRO944.

[0509] In another embodiment, the invention provides isolated PRO944polypeptide. In particular, the invention provides isolated nativesequence PRO944 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 211 of FIG. 98 (SEQ ID NO:270).Additional embodiments of the present invention are directed to PRO944polypeptides comprising amino acids about 22 to 211 of FIG. 98 (SEQ IDNO:270) or amino acid 1 or about 22 to X of FIG. 98 (SEQ ID NO:270),where X is any amino acid from 77 to 86 of FIG. 98 (SEQ ID NO:270).Optionally, the PRO944 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA52185-1370 vector deposited on May 14, 1998 as ATCC 209861.

[0510] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA14007 comprising thenucleotide sequence of FIG. 99 (SEQ ID NO:271).

[0511] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA12733 comprising thenucleotide sequence of FIG. 100 (SEQ ID NO:272).

[0512] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA12746 comprising thenucleotide sequence of FIG. 101 (SEQ ID NO:273).

[0513] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA12834 comprising thenucleotide sequence of FIG. 102 (SEQ ID NO:274).

[0514] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA12846 comprising thenucleotide sequence of FIG. 103 (SEQ ID NO:275).

[0515] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA13104 comprising thenucleotide sequence of FIG. 104 (SEQ ID NO:276).

[0516] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA13259 comprising thenucleotide sequence of FIG. 105 (SEQ ID NO:277).

[0517] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA13959 comprising thenucleotide sequence of FIG. 106 (SEQ ID NO:278).

[0518] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA13961 comprising thenucleotide sequence of FIG. 107 (SEQ ID NO:279).

[0519] 43. PRO983

[0520] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to a vesicle associated protein, VAP-33,wherein the polypeptide is designated in the present application as“PRO983”.

[0521] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO983 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO983polypeptide having amino acid residues 1 to 243 of FIG. 109 (SEQ IDNO:284), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO983 polypeptide having amino acidresidue 1 to X of FIG. 109 (SEQ ID NO:284) where X is any amino acidfrom 219 to 228 of FIG. 109 (SEQ ID NO:284), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA53977-1371 vector deposited on May 14, 1998 as ATCC 209862 whichincludes the nucleotide sequence encoding PRO983.

[0522] In another embodiment, the invention provides isolated PRO983polypeptide. In particular, the invention provides isolated nativesequence PRO983 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 243 of FIG. 109 (SEQ ID NO:284).Additional embodiments of the present invention are directed to PRO983polypeptides comprising amino acid 1 to X of FIG. 109 (SEQ ID NO:284),where Y is any amino acid from 219 to 228 of FIG. 109 (SEQ ID NO:284).Optionally, the PRO983 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA53977-1371 vector deposited on May 14, 1998 as ATCC 209862.

[0523] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA17130 comprising thenucleotide sequence of FIG. 110 (SEQ ID NO:285).

[0524] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA23466 comprising thenucleotide sequence of FIG. 111 (SEQ ID NO:286).

[0525] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA26818 comprising thenucleotide sequence of FIG. 112 (SEQ ID NO:287).

[0526] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA37618 comprising thenucleotide sequence of FIG. 113 (SEQ ID NO:288).

[0527] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA41732 comprising thenucleotide sequence of FIG. 114 (SEQ ID NO:289).

[0528] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA45980 comprising thenucleotide sequence of FIG. 115 (SEQ ID NO:290).

[0529] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA46372 comprising thenucleotide sequence of FIG. 116 (SEQ ID NO:291).

[0530] 44. PRO1057

[0531] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to proteases, wherein the polypeptide isdesignated in the present application as “PRO1057”.

[0532] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1057 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1057polypeptide having amino acid residues 1 to 413 of FIG. 118 (SEQ IDNO:296), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO1057 polypeptide having amino acidresidues about 17 to 413 of FIG. 118 (SEQ ID NO:296), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA57253-1382 vector deposited on May 14, 1998 asATCC 209867 which includes the nucleotide sequence encoding PRO1057.

[0533] In another embodiment, the invention provides isolated PRO1057polypeptide. In particular, the invention provides isolated nativesequence PRO1057 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 413 of FIG. 118 (SEQ ID NO:296).Additional embodiments of the present invention are directed to PRO1057polypeptides comprising amino acids about 17 to 413 of FIG. 118 (SEQ IDNO:296). Optionally, the PRO1057 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA57253-1382 vector deposited on May 14, 1998 as ATCC 209867.

[0534] 45. PRO1071

[0535] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to thrombospondin, wherein the polypeptideis designated in the present application as “PRO1071”.

[0536] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1071 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1071polypeptide having amino acid residues 1 to 525 of FIG. 120 (SEQ IDNO:301), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO1071 polypeptide having amino acidresidues about 26 to 525 of FIG. 120 (SEQ ID NO:301), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA58847-1383 vector deposited on May 20, 1998 asATCC 209879 which includes the nucleotide sequence encoding PRO1071.

[0537] In another embodiment, the invention provides isolated PRO1071polypeptide. In particular, the invention provides isolated nativesequence PRO1071 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 525 of FIG. 120 (SEQ ID NO:301).Additional embodiments of the present invention are directed to PRO1071polypeptides comprising amino acids about 26 to 525 of FIG. 120 (SEQ IDNO:301). Optionally, the PRO1071 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA58847-1383 vector deposited on May 20, 1998 as ATCC 209879.

[0538] 46. PRO1072

[0539] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to reductase proteins, wherein thepolypeptide is designated in the present application as “PRO1072”.

[0540] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1072 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1072polypeptide having amino acid residues 1 to 336 of FIG. 122 (SEQ IDNO:303), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO1072 polypeptide having amino acidresidues about 22 to 336 of FIG. 122 (SEQ ID NO:303), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA58747-1384 vector deposited on May 14, 1998 asATCC 209868 which includes the nucleotide sequence encoding PRO1072.

[0541] In another embodiment, the invention provides isolated PRO1072polypeptide. In particular, the invention provides isolated nativesequence PRO1072 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 336 of FIG. 122 (SEQ ID NO:303).Additional embodiments of the present invention are directed to PRO1072polypeptides comprising amino acids about 22 to 336 of FIG. 122 (SEQ IDNO:303). Optionally, the PRO1072 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA58747-1384 vector deposited on May 14, 1998 as ATCC 209868.

[0542] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA40210 comprising thenucleotide sequence of FIG. 123 (SEQ ID NO:304).

[0543] 47. PRO1075

[0544] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to protein disulfide isomerase, wherein thepolypeptide is designated in the present application as “PRO1075”.

[0545] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1075 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1075polypeptide having amino acid residues 1 to 406 of FIG. 125 (SEQ IDNO:309), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO1075 polypeptide having amino acidresidues about 30 to 406 of FIG. 125 (SEQ ID NO:309), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA57689-1385 vector deposited on May 14, 1998 asATCC 209869 which includes the nucleotide sequence encoding PRO1075.

[0546] In another embodiment, the invention provides isolated PRO1075polypeptide. In particular, the invention provides isolated nativesequence PRO1075 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 406 of FIG. 125 (SEQ ID NO:309).Additional embodiments of the present invention are directed to PRO1075polypeptides comprising amino acids about 30 to 406 of FIG. 125 (SEQ IDNO:309). Optionally, the PRO1075 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA57689-1385 vector deposited on May 14, 1998 as ATCC 209869.

[0547] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA13059 comprising thenucleotide sequence of FIG. 126 (SEQ ID NO:310).

[0548] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA19463 comprising thenucleotide sequence of FIG. 127 (SEQ ID NO:311).

[0549] 48. PRO181

[0550] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to the cornichon protein, wherein thepolypeptide is designated in the present application as “PRO181”.

[0551] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO181 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO181polypeptide having amino acid residues 1 to 144 of FIG. 129 (SEQ IDNO:322), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO181 polypeptide having amino acidresidues about 21 to 144 of FIG. 129 (SEQ ID NO:322) or amino acid 1 orabout 21 to X of FIG. 129 (SEQ ID NO:322) where X is any amino acid from52 to 61 of FIG. 129 (SEQ ID NO:322), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA23330-1390 vector deposited on Apr. 14, 1998 as ATCC 209775 whichincludes the nucleotide sequence encoding PRO181.

[0552] In another embodiment, the invention provides isolated PRO181polypeptide. In particular, the invention provides isolated nativesequence PRO181 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 144 of FIG. 129 (SEQ ID NO:322).Additional embodiments of the present invention are directed to PRO181polypeptides comprising amino acids about 21 to 144 of FIG. 129 (SEQ IDNO:322) or amino acid 1 or about 21 to X of FIG. 129 (SEQ ID NO:322),where X is any amino acid from 52 to 61 of FIG. 129 (SEQ ID NO:322).Optionally, the PRO181 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA23330-1390 vector deposited on Apr. 14, 1998 as ATCC 209775.

[0553] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA13242 comprising thenucleotide sequence of FIG. 130 (SEQ ID NO:323).

[0554] 49. PRO195

[0555] Applicants have identified a cDNA clone that encodes a noveltransmembrane polypeptide, wherein the polypeptide is designated in thepresent application as “PRO195”.

[0556] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO195 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO195polypeptide having amino acid residues 1 to 323 of FIG. 132 (SEQ IDNO:330), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO195 polypeptide having amino acidresidues about 32 to 323 of FIG. 132 (SEQ ID NO:330) or amino acid 1 orabout 32 to X of FIG. 132 (SEQ ID NO:330) where X is any amino acid from236 to 245 of FIG. 132 (SEQ ID NO:330), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA26847-1395 vector deposited on Apr. 14, 1998 as ATCC 209772 whichincludes the nucleotide sequence encoding PRO195.

[0557] In another embodiment, the invention provides isolated PRO195polypeptide. In particular, the ; P invention provides isolated nativesequence PRO195 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 323 of FIG. 132 (SEQ ID NO:330).Additional embodiments of the present invention are directed to PRO195polypeptides comprising amino acids about 32 to 323 of FIG. 132 (SEQ IDNO:330) or amino acid 1 or about 32 to X of FIG. 132 (SEQ ID NO:330),where X is any amino acid from 236 to 245 of FIG. 132 (SEQ ID NO:330).Optionally, the PRO195 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA26847-1395 vector deposited on Apr. 14, 1998 as ATCC 209772.

[0558] In another embodiment, the invention provides an expressedsequence tag (EST) comprising the nucleotide sequence of FIG. 133 (SEQID NO:331), herein designated DNA15062.

[0559] In another embodiment, the invention provides an expressedsequence tag (ESf) comprising the nucleotide sequence of FIG. 134 (SEQID NO:332), herein designated DNA13199.

[0560] 50. PRO865

[0561] Applicants have identified a cDNA clone that encodes a novelsecreted polypeptide, wherein the polypeptide is designated in thepresent application as “PRO865”.

[0562] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO865 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO865polypeptide having amino acid residues 1 to 468 of FIG. 136 (SEQ IDNO:337), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO865 polypeptide having amino acidresidues about 24 to 229 of FIG. 136 (SEQ ID NO:337), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA53974-1401 vector deposited on Apr. 14, 1998as ATCC 209774 which includes the nucleotide sequence encoding PRO865.

[0563] In another embodiment, the invention provides isolated PRO865polypeptide. In particular, the invention provides isolated nativesequence PRO865 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 468 of FIG. 136 (SEQ ID NO:337).An additional embodiment of the present invention is directed to aPRO865 polypeptide comprising amino acids about 24 to 468 of FIG. 136(SEQ ID NO:337). Optionally, the PRO865 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA53974-1401 vector deposited on Apr. 14, 1998 as ATCC 209774.

[0564] In another embodiment, the invention provides an expressedsequence tag (EST) comprising the nucleotide sequence of FIG. 137 (SEQID NO:338), herein designated as DNA37642.

[0565] 51. PRO827

[0566] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to integrin proteins, wherein thepolypeptide is designated in the present application as “PRO827”.

[0567] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO827 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO827polypeptide having amino acid residues 1 to 124 of FIG. 139 (SEQ IDNO:346), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO827 polypeptide having amino acidresidues about 23 to 124 of FIG. 139 (SEQ ID NO:346), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA57039-1402 vector deposited on Apr. 14, 1998as ATCC 209777 which includes the nucleotide sequence encoding PRO827.

[0568] In another embodiment, the invention provides isolated PRO827polypeptide. In particular, the invention provides isolated nativesequence PRO827 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 124 of FIG. 139 (SEQ ID NO:346).An additional embodiment of the present invention is directed to aPRO827 polypeptide comprising amino acids about 23 to 124 of FIG. 139(SEQ ID NO:346). Optionally, the PRO827 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe DNA57039-1402 vector deposited on Apr. 14, 1998 as ATCC 209777.

[0569] 52. PRO1114

[0570] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to cytoline receptor family4 proteins,wherein the polypeptide is designated in the present application as“PRO1114”.

[0571] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1114 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1114polypeptide having amino acid residues 1 to 311 of FIG. 142 (SEQ IDNO:352), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO1114 polypeptide having amino acidresidues about 30 to 311 of FIG. 142 (SEQ ID NO:352) or amino acid 1 orabout 30 to X of FIG. 142 (SEQ ID NO:352), where X is any amino acidfrom 225 to 234 of FIG. 142 (SEQ ID NO:352), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA57033-1403 vector deposited on May 27, 1998 as ATCC 209905 whichincludes the nucleotide sequence encoding PRO1114.

[0572] In another embodiment, the invention provides isolated PRO1114polypeptide. In particular, the invention provides isolated nativesequence PRO1114 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 311 of FIG. 142 (SEQ ID NO:352).Additional embodiments of the present invention are directed to PRO114polypeptides comprising amino acids about 30 to 311 of FIG. 142 (SEQ IDNO:352) or amino acid 1 or about 30 to X of FIG. 142 (SEQ ID NO:352),where X is any amino acid from 225 to 234 of FIG. 142 (SEQ ID NO:352).Optionally, the PRO1114 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA57033-1403 vector deposited on May 27, 1998 as ATCC 209905.

[0573] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA48466 comprising thenucleotide sequence of FIG. 143 (SEQ ID NO:353).

[0574] A cDNA clone (DNA57033-1403) has been identified that encodes anovel interferon receptor polypeptide, designated in the presentapplication as “PRO1114 interferon receptor”.

[0575] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1114 interferon receptorpolypeptide.

[0576] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO1114 interferon receptor polypeptide having thesequence of amino acid residues from about 1 or about 30 to about 311,inclusive of FIG. 142 (SEQ ID NO:352), or (b) the complement of the DNAmolecule of (a).

[0577] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO1114 interferon receptor polypeptidecomprising DNA hybridizing to the complement of the nucleic acid betweenabout nucleotides 250 or about 337 and about 1182, inclusive, of FIG.141 (SEQ ID NO:351). Preferably, hybridization occurs under stringenthybridization and wash conditions.

[0578] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 209905 (DNA57033-1403) or (b) the complement of the nucleicacid molecule of (a). In a preferred embodiment, the nucleic acidcomprises a DNA encoding the same mature polypeptide encoded by thehuman protein cDNA in ATCC Deposit No. 209905 (DNA57033-1403).

[0579] In still a further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues 1 or about 30 to about 311, inclusive of FIG. 142(SEQ ID NO:352), or (b) the complement of the DNA of (a).

[0580] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least 10 nucleotides and produced by hybridizinga test DNA molecule under stringent conditions with (a) a DNA moleculeencoding a PRO1114 interferon receptor polypeptide having the sequenceof amino acid residues from 1 or about 30 to about 311, inclusive ofFIG. 142 (SEQ ID NO:352), or (b) the complement of the DNA molecule of(a), and, if the DNA molecule has at least about an 80% sequenceidentity, prefereably at least about an 85% sequence identity, morepreferably at least about a 90% sequence identity, most preferably atleast about a 95% sequence identity to (a) or (b), isolating the testDNA molecule.

[0581] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1114 interferon receptorpolypeptide, with or without the N-terminal signal sequence and/or theinitiating methionine, and its soluble, i.e., transmembrane domaindeleted or inactivated variants, or is complementary to such encodingnucleic acid molecule. The signal peptide has been tentativelyidentified as extending from about amino acid position 1 to about aminoacid position 29 in the sequence of FIG. 142 (SEQ ID NO:352). Thetransmembrane domain has been tentatively identified as extending fromabout amino acid position 230 to about amino acid position 255 in thePRO114 interferon receptor amino acid sequence (FIG. 142, SEQ IDNO:352).

[0582] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 1or about 30 to about 311, inclusive of FIG. 142 (SEQ ID NO:352), or (b)the complement of the DNA of (a).

[0583] Another embodiment is directed to fragments of a PRO1114interferon receptor polypeptide coding sequence that may find use ashybridization probes. Such nucleic acid fragments are from about 20 toabout 80 nucleotides in length, preferably from about 20 to about 60nucleotides in length, more preferably from about 20 to about 50nucleotides in length and most preferably from about 20 to about 40nucleotides in length and may be derived from the nucleotide sequenceshown in FIG. 141 (SEQ ID NO:351).

[0584] In another embodiment, the invention provides a vector comprisingDNA encoding PRO1114 interferon receptor or its variants. The vector maycomprise any of the isolated nucleic acid molecules hereinaboveidentified.

[0585] A host cell comprising such a vector is also provided. By way ofexample, the host cells may be CHO cells, E. coli, or yeast. A processfor producing PRO1114 interferon receptor polypeptides is furtherprovided and comprises culturing host cells under conditions suitablefor expression of PRO1114 interferon receptor and recovering PRO1114interferon receptor from the cell culture.

[0586] In another embodiment, the invention provides isolated PRO1114interferon receptor polypeptide encoded by any of the isolated nucleicacid sequences hereinabove identified.

[0587] In a specific aspect, the invention provides isolated nativesequence PRO1114 interferon receptor polypeptide, which in certainembodiments, includes an amino acid sequence comprising residues 1 orabout 30 to about 311 of FIG. 142 (SEQ ID NO:352).

[0588] In another aspect, the invention concerns an isolated PRO1114interferon receptor polypeptide, comprising an amino acid sequencehaving at least about 80% sequence identity, preferably at least about85% sequence identity, more preferably at least about 90% sequenceidentity, most preferably at least about 95% sequence identity to thesequence of amino acid residues 1 or about 30 to about 311, inclusive ofFIG. 142 (SEQ ID NO:352).

[0589] In a further aspect, the invention concerns an isolated PRO1114interferon receptor polypeptide, comprising an amino acid sequencescoring at least about 80% positives, preferably at least about 85%positives, more preferably at least about 90% positives, most preferablyat least about 95% positives when compared with the amino acid sequenceof residues 1 or about 30 to about 311, inclusive of FIG. 142 (SEQ IDNO:352).

[0590] In yet another aspect, the invention concerns an isolated PRO1114interferon receptor polypeptide, comprising the sequence of amino acidresidues 1 or about 30 to about 311, inclusive of FIG. 142 (SEQ IDNO:352), or a fragment thereof sufficient to provide a binding site foran anti-PRO1114 interferon receptor antibody. Preferably, the PRO1114interferon receptor fragment retains a qualitative biological activityof a native PRO1114 interferon receptor polypeptide.

[0591] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO1114 interferonreceptor polypeptide having the sequence of amino acid residues fromabout 1 or about 30 to about 311, inclusive of FIG. 142 (SEQ ID NO:352),or (b) the complement of the DNA molecule of (a), and if the test DNAmolecule has at least about an 80% sequence identity, preferably atleast about an 85% sequence identity, more preferably at least about a90% sequence identity, most preferably at least about a 95% sequenceidentity to (a) or (b), (ii) culturing a host cell comprising the testDNA molecule under conditions suitable for expression of thepolypeptide, and (iii) recovering the polypeptide from the cell culture.

[0592] In another embodiment, the invention provides chimeric moleculescomprising a PRO1114 interferon receptor polypeptide fused to aheterologous polypeptide or amino acid sequence. An example of such achimeric molecule comprises a PRO114 interferon receptor polypeptidefused to an epitope tag sequence or a Fc region of an immunoglobulin.

[0593] In another embodiment, the invention provides an antibody whichspecifically binds to a PRO1114 interferon receptor polypeptide.Optionally, the antibody is a monoclonal antibody.

[0594] In yet another embodiment, the invention concerns agonists andantagonists of a native PRO1114 interferon receptor polypeptide. In aparticular embodiment, the agonist or antagonist is an anti-PRO1114interferon receptor antibody.

[0595] In a further embodiment, the invention concerns a method ofidentifying agonists or antagonists of a native PRO1114 interferonreceptor polypeptide by contacting the native PRO1114 interferonreceptor polypeptide with a candidate molecule and monitoring abiological activity mediated by said polypeptide.

[0596] In a still further embodiment, the invention concerns acomposition comprising a PRO1114 interferon receptor polypeptide, or anagonist or antagonist as hereinabove defined, in combination with apharmaceutically acceptable carrier.

[0597] 53. PRO237

[0598] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to carbonic anhydrase, wherein thepolypeptide is designated in the present application as “PRO237”.

[0599] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO237 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO237polypeptide having amino acid residues 1 to 328 of FIG. 145 (SEQ IDNO:358), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO237 polypeptide having amino acidresidues about 24 to 328 of FIG. 145 (SEQ ID NO:358) or amino acid 1 orabout 24 to X of FIG. 145 (SEQ ID NO:358), where X is any amino acidfrom 172 to 181 of FIG. 145 (SEQ ID NO:358), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA34353-1428 vector deposited on May 12, 1998 as ATCC 209855 whichincludes the nucleotide sequence encoding PRO237.

[0600] In another embodiment, the invention provides isolated PRO237polypeptide. In particular, the invention provides isolated nativesequence PRO237 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 328 of FIG. 145 (SEQ ID NO:358).Additional embodiments of the present invention are directed to PRO237polypeptides comprising amino acids about 24 to 328 of FIG. 145 (SEQ IDNO:358) or amino acid 1 or about 24 to X of FIG. 145 (SEQ ID NO:358),where X is any amino acid from 172 to 181 of FIG. 145 (SEQ ID NO:358).Optionally, the PRO237 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA34353-1428 vector deposited on May 12, 1998 as ATCC 209855.

[0601] 54. PRO541

[0602] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to a trypsin inhibitor protein, wherein thepolypeptide is designated in the present application as “PRO541”.

[0603] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO541 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO541polypeptide having amino acid residues 1 to 500 of FIG. 147 (SEQ IDNO:363), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO541 polypeptide having amino acidresidues about 21 to 500 of FIG. 147 (SEQ ID NO:363), or iscomplementary to such encoding nucleic acid sequence, and remains stablybound to it under at least moderate, and optionally, under highstringency conditions. The isolated nucleic acid sequence may comprisethe cDNA insert of the DNA45417-1432 vector deposited on May 27, 1998 asATCC 209910 which includes the nucleotide sequence encoding PRO541.

[0604] In another embodiment, the invention provides isolated PRO541polypeptide. In particular, the invention provides isolated nativesequence PRO541 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 500 of FIG. 147 (SEQ ID NO:363).Additional embodiments of the present invention are directed to PRO541polypeptides comprising amino acids about 21 to 500 of FIG. 147 (SEQ IDNO:363). Optionally, the PRO541 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of theDNA45417-1432 vector deposited on May 27, 1998 as ATCC 209910.

[0605] 55. PRO273

[0606] Applicants have identified a cDNA clone that encodes a novelpolypeptide, wherein the polypeptide is designated in the presentapplication as “PRO273”.

[0607] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO273 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO273polypeptide having amino acid residues 1 through 111 of FIG. 149 (SEQ IDNO:370), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In another embodiment, the inventionprovides isolated PRO273 polypeptide. In particular, the inventionprovides isolated native sequence PRO273 polypeptide, which in oneembodiment, includes an amino acid sequence comprising residues 1through 111 of FIG. 149 (SEQ ID NO:370).

[0608] 56. PRO701

[0609] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to neuroligins 1, 2, and 3, wherein thepolypeptide is designated in the present application as “PRO701”.

[0610] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO701 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO701polypeptide having amino acid residues 1 through 816 of FIG. 151 (SEQ IDNO:375), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited with the ATCC on Mar.31, 1998 which includes the nucleotide sequence encoding PRO701.

[0611] In another embodiment, the invention provides isolated PRO701polypeptide. In particular, the invention provides isolated nativesequence PRO701 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 816 of FIG. 151 (SEQ IDNO:375). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO701 polypeptide. Optionally,the PRO701 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited with theATCC on Mar. 31, 1998.

[0612] 57. PRO704

[0613] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with VIP36, wherein the polypeptideis designated in the present application as “PRO704”.

[0614] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO704 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO704polypeptide having amino acid residues 1 through 348 of FIG. 153 (SEQ IDNO:380), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Mar. 31, 1998 withthe ATCC as DNA50911-1288, which includes the nucleotide sequenceencoding PRO704.

[0615] In another embodiment, the invention provides isolated PRO704polypeptide. In particular, the invention provides isolated nativesequence PRO704 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 348 of FIG. 153 (SEQ IDNO:380). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO704 polypeptide. Optionally,the PRO704 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Mar.31, 1998 with the ATCC as DNA50911-1288.

[0616] 58. PRO706

[0617] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to prostatic acid phosphatase precursor andlysosomal acid phosphatase precursor, wherein the polypeptide isdesignated in the present application as “PRO706”.

[0618] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO706 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO706polypeptide having amino acid residues 1 through 480 of FIG. 155 (SEQ IDNO:385), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Apr. 21, 1998 withthe ATCC as DNA48329-1290 which includes the nucleotide sequenceencoding PRO706.

[0619] In another embodiment, the invention provides isolated PRO706polypeptide. In particular, the invention provides isolated nativesequence PRO706 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 480 of FIG. 155 (SEQ IDNO:385), or comprising residues 19 through 480 of FIG. 155 (SEQ IDNO:385). Optionally, the PRO706 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of the vectordeposited on Apr. 21, 1998 with the ATCC as DNA48329-1290.

[0620] 59. PRO707

[0621] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to cadherins, particularly cadherin FIB3,wherein the polypeptide is designated in the present application as“PRO707”.

[0622] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO707 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO707polypeptide having amino acid residues 1 to 916 of FIG. 157 (SEQ IDNO:390), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 27, 1998 withthe ATCC as DNA48306-1291 which includes the nucleotide sequenceencoding PRO707.

[0623] In another embodiment, the invention provides isolated PRO707polypeptide. In particular, the invention provides isolated nativesequence PRO707 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 916 of FIG. 157 (SEQ ID NO:390).An additional embodiment of the present invention is directed to anisolated extracellular domain of a PRO707 polypeptide. Optionally, thePRO707 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on May27, 1998 with the ATCC as DNA48306-1291.

[0624] 60. PRO322

[0625] Applicants have identified a cDNA clone that encodes a novelpolypeptide having homology to neuropsin, wherein the polypeptide isdesignated in the present application as “PRO322”.

[0626] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO322 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO322polypeptide having amino acid residues 1 or 24 through 260 of FIG. 159(SEQ ID NO:395), or is complementary to such encoding nucleic acidsequence, and remains stably bound to it under at least moderate, andoptionally, under high stringency conditions. The isolated nucleic acidsequence may comprise the cDNA insert of the vector deposited on Mar.11, 1998 as ATCC no. 209669 which includes the nucleotide sequenceencoding PRO322.

[0627] In another embodiment, the invention provides isolated PRO322polypeptide. In particular, the invention provides isolated nativesequence PRO322 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 or 24 through 260 of FIG. 159 (SEQID NO:395). An additional embodiment of the present invention isdirected to an isolated extracellular domain of a PRO322 polypeptide.Optionally, the PRO322 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of the vectordeposited on Mar. 11, 1998 as ATCC no. 209669.

[0628] 61. PRO526

[0629] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with ALS, wherein the polypeptideis designated in the present application as “PRO526”.

[0630] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO526 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO526polypeptide having amino acid residues 1 to 473 of FIG. 161 (SEQ IDNO:400), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Mar. 26, 1998 withthe ATCC as DNA44184-1319 which includes the nucleotide sequenceencoding PRO526.

[0631] In another embodiment, the invention provides isolated PRO526polypeptide. In particular, the invention provides isolated nativesequence PRO526 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 473 of FIG. 161 (SEQ ID NO:400).Optionally, the PRO526 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of the vectordeposited on Mar. 26, 1998 with the ATCC as DNA44184-1319 which includesthe nucleotide sequence encoding PRO526.

[0632] 62. PRO531

[0633] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with protocadherins, wherein thepolypeptide is designated in the present application as “PRO531 ”.

[0634] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO531 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO531polypeptide having amino acid residues 1 to 789 of FIG. 163 (SEQ IDNO:405), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Mar. 26, 1998 asDNA48314-1320 which includes the nucleotide sequence encoding PRO531.

[0635] In another embodiment, the invention provides isolated PRO531polypeptide. In particular, the invention provides isolated nativesequence PRO531 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 789 of FIG. 163 (SEQ ID NO:405).An additional embodiment of the present invention is directed to anisolated extracellular domain of a PRO531 polypeptide. Optionally, thePRO531 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Mar.26, 1998 as DNA48314-1320.

[0636] 63. PRO534

[0637] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with disulfide isomerase (sometimesreferred to herein as protein disulfide isomerase), wherein thepolypeptide is designated in the present application as “PRO534”.

[0638] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO534 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO534polypeptide having amino acid residues 1 to 360 of FIG. 165 (SEQ TDNO:410), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Mar. 26, 1998 asDNA48333-1321 which includes the nucleotide sequence encoding PRO534.

[0639] In another embodiment, the invention provides isolated PRO534polypeptide. In particular, the invention provides isolated nativesequence PRO534 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 360 of FIG. 165 (SEQ ID NO:410).An additional embodiment of the present invention is directed to anisolated extracellular domain of a PRO534 polypeptide. Optionally, thePRO534 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Mar.26, 1998 as DNA48333-1321.

[0640] 64. PRO697

[0641] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with sFRPs, wherein the polypeptideis designated in the present application as “PRO697”.

[0642] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO697 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO697polypeptide having amino acid residues 1 through 295 of FIG. 167 (SEQ IDNO:415), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited with the ATCC on Mar.26, 1998 as DNA50920-1325 which includes the nucleotide sequenceencoding PRO697.

[0643] In another embodiment, the invention provides isolated PRO697polypeptide. In particular, the invention provides isolated nativesequence PRO697 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 295 of FIG. 167 (SEQ IDNO:415). Optionally, the PRO697 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of the vectordeposited with the ATCC on Mar. 26, 1998 as DNA50920-1325.

[0644] 65. PRO717

[0645] Applicants have identified a cDNA clone that encodes a novel 12transmembrane polypeptide, wherein the polypeptide is designated in thepresent application as “PRO717”.

[0646] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO717 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO717polypeptide having amino acid residues 1 through 560 of FIG. 169 (SEQ IDNO:420), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Apr. 28, 1998 withthe ATCC as DNA50988-1326 which includes the nucleotide sequenceencoding PRO717.

[0647] In another embodiment, the invention provides isolated PRO717polypeptide. In particular, the invention provides isolated nativesequence PRO717 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 560 of FIG. 169 (SEQ IDNO:420). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO717 polypeptide. Optionally,the PRO717 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Apr.28, 1998 with the ATCC as DNA50988-1326.

[0648] 66. PRO731

[0649] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with protocadherin 4, wherein thepolypeptide is designated in the present application as “PRO731”.

[0650] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO731 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO731polypeptide having amino acid residues 1 through 1184 of FIG. 171 (SEQID NO:425), or is complementary to such encoding nucleic acid sequence,and remains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Mar. 31, 1998 withthe ATCC as DNA48331-1329 which includes the nucleotide sequenceencoding PRO731.

[0651] In another embodiment, the invention provides isolated PRO731polypeptide. In particular, the invention provides isolated nativesequence PRO731 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 1184 of FIG. 171 (SEQ IDNO:425). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO731 polypeptide. Optionally,the PRO731 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Mar.31, 1998 with the ATCC as DNA48331-1329.

[0652] 67. PRO218

[0653] Applicants have identified a cDNA clone that encodes a novelmulti-transmembrane protein having sequence identity with membraneregulator proteins, wherein the polypeptide is designated in the presentapplication as “PRO218”.

[0654] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO218 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO218polypeptide having amino acid residues 1 through 455 of FIG. 173 (SEQ IDNO:430), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Apr. 28, 1998 withthe ATCC as DNA30867-1335 which includes the nucleotide sequenceencoding PRO218.

[0655] In another embodiment, the invention provides isolated PRO218polypeptide. In particular, the invention provides isolated nativesequence PRO218 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 455 of FIG. 173 (SEQ IDNO:430). Optionally, the PRO218 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of the vectordeposited on Apr. 28, 1998 with the ATCC as DNA30867-1335.

[0656] In another embodiment, the invention provides an expressedsequence tag (EST) sequence comprising the nucleotide sequence of FIG.174 (SEQ ID NO:431), designated herein as DNA14472.

[0657] In another embodiment, the invention provides an expressedsequence tag (EST) sequence comprising the nucleotide sequence of FIG.175 (SEQ ID NO:432), designated herein as DNA15846.

[0658] 68. PRO768

[0659] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with integrins, wherein thepolypeptide is designated in the present application as “PRO768”.

[0660] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO768 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO768polypeptide having amino acid residues 1 through 1141 of FIG. 177 (SEQID NO:437), or is complementary to such encoding nucleic acid sequence,and remains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Apr. 6, 1998 asDNA55737-1345 which includes the nucleotide sequence encoding PRO768.

[0661] In another embodiment, the invention provides isolated PRO768polypeptide. In particular, the invention provides isolated nativesequence PRO768 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 1141 of FIG. 177 (SEQ IDNO:437). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO768 polypeptide. Optionally,the PRO768 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Apr.6, 1998 as DNA55737-1345.

[0662] 69. PRO771

[0663] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with testican, wherein thepolypeptide is designated in the present application as “PRO771”.

[0664] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO771 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO771polypeptide having amino acid residues 1 through 436 of FIG. 179 (SEQ IDNO:442), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Apr. 7, 1998 withthe ATCC as DNA49829-1346 which includes the nucleotide sequenceencoding PRO771.

[0665] In another embodiment, the invention provides isolated PRO771polypeptide. In particular, the invention provides isolated nativesequence PRO771 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 436 of FIG. 179 (SEQ IDNO:442). Optionally, the PRO771 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of the vectordeposited on Apr. 7, 1998 with the ATCC as DNA49829-1346.

[0666] 70. PRO733

[0667] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with the T1I/ST2 receptor bindingprotein, wherein the polypeptide is designated in the presentapplication as “PRO733”.

[0668] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO733 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO733polypeptide having amino acid residues 1 through 229 of FIG. 181 (SEQ IDNO:447), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on Apr. 7, 1998 withthe ATCC as DNA52196-1348 which includes the nucleotide sequenceencoding PRO733.

[0669] In another embodiment, the invention provides isolated PRO733polypeptide. In particular, the invention provides isolated nativesequence PRO733 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 229 of FIG. 181 (SEQ IDNO:447). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO733 polypeptide. Optionally,the PRO733 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on Apr.7, 1998 as DNA52196-1348.

[0670] 71. PRO162

[0671] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with pancreatitis-associatedprotein, wherein the polypeptide is designated in the presentapplication as “PRO162”.

[0672] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO162 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO162polypeptide having amino acid residues 1 through 175 of FIG. 183 (SEQ IDNO:452), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 6, 1998 with theATCC as DNA56965-1356 which includes the nucleotide sequence encodingPRO162.

[0673] In another embodiment, the invention provides isolated PRO162polypeptide. In particular, the invention provides isolated nativesequence PRO162 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 175 of FIG. 183 (SEQ IDNO:452). Optionally, the PRO162 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of the vectordeposited on May 6, 1998 with the ATCC as DNA56965-1356.

[0674] 72. PRO788

[0675] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with anti-neoplastic urinaryprotein, wherein the polypeptide is designated in the presentapplication as “PRO788”.

[0676] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO788 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO788polypeptide having amino acid residues 1 through 125 of FIG. 185 (SEQ IDNO:454), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 6, 1998 with theATCC as DNA56405-1357 which includes the nucleotide sequence encodingPRO788.

[0677] In another embodiment, the invention provides isolated PRO788polypeptide. In particular, the invention provides isolated nativesequence PRO788 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 125 of FIG. 185 (SEQ IDNO:454). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO788 polypeptide. Optionally,the PRO788 polypeptide is obtained or is obtainable by expressing thepolypeptide encoded by the cDNA insert of the vector deposited on May 6,1998 with the ATCC as DNA56405-1357.

[0678] 73. PRO1008

[0679] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with dickkopf-1 (dkk-1), whereinthe polypeptide is designated in the present application as “PRO1008”.

[0680] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1008 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1008polypeptide having amino acid residues 1 through 266 of FIG. 187 (SEQ IDNO:456), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 20, 1998 withthe ATCC as DNA57530-1375 which includes the nucleotide sequenceencoding PRO1008.

[0681] In another embodiment, the invention provides isolated PRO1008polypeptide. In particular, the invention provides isolated nativesequence PRO1008 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 266 of FIG. 187 (SEQ IDNO:456). Optionally, the PRO1008 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe vector deposited on May 20, 1998 with the ATCC as DNA57530-1375.

[0682] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA16508 comprising thenucleotide sequence of FIG. 188 (SEQ ID NO:457).

[0683] 74. PRO1012

[0684] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with disulfide isomerase andphospholipase C, wherein the polypeptide is designated in the presentapplication as “PRO1012”.

[0685] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1012 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1012polypeptide having amino acid residues 1 through 747 of FIG. 190 (SEQ IDNO:459), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 14, 1998 withthe ATCC as DNA56439-1376, which includes the nucleotide sequenceencoding PRO1012.

[0686] In another embodiment, the invention provides isolated PRO1012polypeptide. In particular, the invention provides isolated nativesequence PRO1012 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 747 of FIG. 190 (SEQ IDNO:459). Optionally, the PRO1012 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe vector deposited on May 14, 1998 with the ATCC as DNA56439-1376.

[0687] 75. PRO1014

[0688] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with reductase, wherein thepolypeptide is designated in the present application as “PRO1014”.

[0689] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1014 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1014polypeptide having amino acid residues 1 through 300 of FIG. 192 (SEQ IDNO:464), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 20, 1998 asDNA56409-1377 with the ATCC which includes the nucleotide sequenceencoding PRO1014.

[0690] In another embodiment, the invention provides isolated PRO1014polypeptide. In particular, the invention provides isolated nativesequence PRO1014 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 300 of FIG. 192 (SEQ IDNO:464). Optionally, the PRO1014 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe vector deposited on May 20, 1998 as DNA56409-1377 with the ATCC.

[0691] 76. PRO1017

[0692] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with HNK-1 sulfotransferase,wherein the polypeptide is designated in the present application as“PRO1017”.

[0693] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1017 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1017polypeptide having amino acid residues 1 through 414 of FIG. 194 (SEQ IDNO:466), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 20, 1998 withthe ATCC as DNA56112-1379 which includes the nucleotide sequenceencoding PRO1017.

[0694] In another embodiment, the invention provides isolated PRO1017polypeptide. In particular, the invention provides isolated nativesequence PRO1017 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 414 of FIG. 194 (SEQ IDNO:466). Optionally, the PRO1017 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe vector deposited on May 20, 1998 with the ATCC as DNA56112-1379.

[0695] 77. PRO474

[0696] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with dehydrogenase, wherein thepolypeptide is designated in the present application as “PRO474”.

[0697] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO474 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO474polypeptide having amino acid residues 1 through 270 of FIG. 196 (SEQ IDNO:468), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 14, 1998 withthe ATCC as DNA56045-1380 which includes the nucleotide sequenceencoding PRO474.

[0698] In another embodiment, the invention provides isolated PRO474polypeptide. In particular, the invention provides isolated nativesequence PRO474 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 270 of FIG. 196 (SEQ IDNO:468). Optionally, the PRO474 polypeptide is obtained or is obtainableby expressing the polypeptide encoded by the cDNA insert of the vectordeposited on May 14, 1998 with the ATCC as DNA56045-1380.

[0699] 78. PRO1031

[0700] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with IL-17, wherein the polypeptideis designated in the present application as “PRO1031”.

[0701] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1031 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1031polypeptide having amino acid residues 1 through 180 of FIG. 198 (SEQ IDNO:470), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 14, 1998 withthe ATCC as DNA59294-1381 which includes the nucleotide sequenceencoding PRO1031.

[0702] In another embodiment, the invention provides isolated PRO1031polypeptide. In particular, the invention provides isolated nativesequence PRO1031 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 180 of FIG. 198 (SEQ IDNO:470). Optionally, the PRO1031 polypeptide is obtained or isobtainable by expressing the polypeptide encoded by the cDNA insert ofthe vector deposited on May 14, 1998 with the ATCC as DNA59294-1381.

[0703] 79. PRO938

[0704] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity to protein disulfide isomerase,wherein the polypeptide is designated in the present application as“PRO938”.

[0705] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO938 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO938polypeptide having amino acid residues 1 to 349 of FIG. 200 (SEQ IDNO:472), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. In other aspects, the isolated nucleicacid comprises DNA encoding the PRO938 polypeptide having amino acidresidues about 23 to 349 of FIG. 200 (SEQ ID NO:472) or amino acid 1 orabout 23 to X of FIG. 200 (SEQ ID NO:472), where X is any amino acidfrom 186 to 195 of FIG. 200 (SEQ ID NO:472), or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of theDNA56433-1406 vector deposited on May 12, 1998, as ATCC Accession No.209857 which includes the nucleotide sequence encoding PRO938.

[0706] In another embodiment, the invention provides isolated PRO938polypeptide. In particular, the invention provides isolated nativesequence PRO938 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 349 of FIG. 200 (SEQ ID NO:472).Additional embodiments of the present invention are directed to PRO938polypeptides comprising amino acids about 23 to 349 of FIG. 200 (SEQ IDNO:472) or amino acid 1 or about 23 to X of FIG. 200 (SEQ ID NO:472),where X is any amino acid from 186 to 195 of FIG. 200 (SEQ ID NO:472).Optionally, the PRO938 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of theDNA56433-1406 vector deposited on May 12, 1998, as ATCC Accession No.209857.

[0707] 80. PRO1082

[0708] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with a lectin-like oxidized LDLreceptor, wherein the polypeptide is designated in the presentapplication as “PRO1082”.

[0709] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1082 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1082polypeptide having amino acid residues 1 through 201 of FIG. 202 (SEQ IDNO:477), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 14, 1998 withthe ATCC as DNA53912-1457 which includes the nucleotide sequenceencoding PRO1082.

[0710] In another embodiment, the invention provides isolated PRO1082polypeptide. In particular, the invention provides isolated nativesequence PRO1082 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 201 of FIG. 202 (SEQ IDNO:477). An additional embodiment of the present invention ix directedto an isolated domain of a PRO1082 polypeptide, excluding thetransmembrane domain. Optionally, the PRO1082 polypeptide is obtained oris obtainable by expressing the polypeptide encoded by the cDNA insertof the vector deposited on May 14, 1998 with the ATCC as DNA53912-1457.

[0711] 81. PRO1083

[0712] Applicants have identified a cDNA clone that encodes a novelpolypeptide having sequence identity with a 7TM receptor,latrophilin-related protein 1, and a macrophage restricted cell surfaceglycoprotein, wherein the polypeptide is designated in the presentapplication as “PRO1083”.

[0713] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1083 polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the PRO1083polypeptide having amino acid residues 1 through 693 of FIG. 204 (SEQ IDNO:483), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions. The isolated nucleic acid sequence maycomprise the cDNA insert of the vector deposited on May 12, 1998 withthe ATCC as DNA50921-1458 which includes the nucleotide sequenceencoding PRO1083.

[0714] In another embodiment, the invention provides isolated PRO1083polypeptide. ln particular, the invention provides isolated nativesequence PRO1083 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 693 of FIG. 204 (SEQ IDNO:483). An additional embodiment of the present invention is directedto an isolated extracellular domain of a PRO1083 polypeptide.Optionally, the PRO1083 polypeptide is obtained or is obtainable byexpressing the polypeptide encoded by the cDNA insert of the vectordeposited on May 12, 1998 with the ATCC as DNA50921-1458.

[0715] In another embodiment, the invention provides an expressedsequence tag (EST) designated herein as DNA24256 which comprises thenucleotide sequence of FIG. 205 (SEQ ID NO:484).

[0716] 82. PRO200

[0717] The objects of this invention, as defined generally supra, areachieved at least in part by the provision of a novel polypeptide,VEGF-E also herein designated PRO200, (SEQ ID NO:488) and the nucleicacid encoding therefor, SEQ ID NO:487, residues 259 through 1293.

[0718] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a VEGF-E polypeptide. In oneaspect, the isolated nucleic acid comprises DNA encoding the VEGF-Epolypeptide having amino acid residues 1 through 345 of FIG. 207 (SEQ IDNO:488), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under low stringency conditions. In anotherembodiment, variants are provided wherein the VEGF-E nucleic acid hassingle or multiple deletions, substitutions, insertions, truncations orcombinations thereof.

[0719] In another embodiment, the invention provides isolated VEGF-Epolypeptide. In particular, the invention provides an isolated nativesequence VEGF-E polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 through 345 of FIG. 207 (SEQ IDNO:488). In another embodiment, variants are provided wherein the VEGF-Epolypeptide has single or multiple deletions, substitutions, insertions,truncations or combinations thereof.

[0720] In yet further embodiments, the present invention is directed tocompositions useful for treating indications where proliferation,survival and/or differentiation of cells is desired, comprising atherapeutically effective amount of a VEGF-E polypeptide hereof inadmixture with a pharmaceutically acceptable carrier.

[0721] The invention further includes associated embodiments of VEGF-Esuch as modified VEGF-E polypeptides and modified variants which havethe same biological applications as VEGF-E, and pharmaceuticalcompositions incorporating same. Inhibitors of VEGF-E are also provided.

[0722] 83. PRO285 and PRO286

[0723] Applicants have identified two novel cDNA clones that encodenovel human Toll polypeptides, designated in the present application asPRO285 (encoded by DNA40021-1154) and PRO286 (encoded by DNA42663-1154).

[0724] In one embodiment, the invention provides an isolated nucleicacid molecule comprising a DNA encoding a polypeptide having at leastabout 80% sequence identity, preferably at least about 85% sequenceidentity, more preferably at least about 90% sequence identity, mostpreferably at least about 95% sequence identity to (a) a DNA moleculeencoding a PRO285 polypeptide having amino acid residues 27 to 839 ofFIG. 209 (SEQ ID NO:496); or (b) to a DNA molecule encoding a PRO286polypeptide having amino acid residues 27 to 825 of FIG. 211 (SEQ IDNO:498) or (c) the complement of the DNA molecule of (a) or (b). Thecomplementary DNA molecule preferably remains stably bound to suchencoding nucleic acid sequence under at least moderate, and optionally,under high stringency conditions.

[0725] In a further embodiment, the isolated nucleic acid moleculecomprises a polynucleotide that has at least about 90%, preferably atleast about 95% sequence identity with a polynucleotide encoding apolypeptide comprising the sequence of amino acids 1 to 839 of FIG. 209(SEQ ID NO:496); or at least about 90%, preferably at least about 95%sequence identity with a polynucleotide encoding a polypeptidecomprising the sequence of amino acids 1 to 1041 of FIG. 211 (SEQ IDNO:498).

[0726] In a specific embodiment, the invention provides an isolatednucleic acid molecule comprising DNA encoding native or variant PRO285and PRO286 polypeptides, with or without the N-terminal signal sequence,and with or without the transmembrane regions of the respectivefull-length sequences. In one aspect, the isolated nucleic acidcomprises DNA encoding a mature, full-length native PRO285 or PRO286polypeptide having amino acid residues 1 to 1049 of FIG. 209 (SEQ IDNO:496) and 1 to 1041 of FIG. 211 (SEQ ID NO:498), or is complementaryto such encoding nucleic acid sequence. In another aspect, the inventionconcerns an isolated nucleic acid molecule that comprises DNA encoding anative PRO285 or PRO286 polypeptide without an N-terminal signalsequence, or is complementary to such encoding nucleic acid sequence. Inyet another embodiment, the invention concerns nucleic acid encodingtransmembrane-domain deleted or inactivated forms of the full-lengthnative PRO285 or PRO286 proteins.

[0727] In another embodiment, the invention the isolated nucleic acidmolecule comprises the clone (DNA40021-1154) deposited on Oct. 17, 1997,under ATCC number 209389; or the clone (DNA42663-1154) deposited on Oct.17, 1997, under ATCC number 209386.

[0728] In yet another embodiment, the invention provides a vectorcomprising DNA encoding PRO285 and PRO286 polypeptides, or theirvariants. Thus, the vector may comprise any of the isolated nucleic acidmolecules hereinabove defined.

[0729] In another embodiment, the invention provides isolated PRO285 andPRO286 polypeptides. In particular, the invention provides isolatednative sequence PRO285 and PRO286 polypeptides, which in one embodiment,include the amino acid sequences comprising residues 1 to 1049 and 1 to1041 of FIGS. 209 and 211 (SEQ ID NOS:496 and 498), respectively. Theinvention also provides for variants of the PRO285 and PRO286polypeptides which are encoded by any of the isolated nucleic acidmolecules hereinabove defined. Specific variants include, but are notlimited to, deletion (truncated) variants of the full-length nativesequence PRO285 and PRO286 polypeptides which lack the respectiveN-terminal signal sequences and/or have their respective transmembraneand/or cytoplasmic domains deleted or inactivated.

[0730] The invention also specifically includes antibodies with dualspecificities, e.g., bispecific antibodies binding more than one Tollpolypeptide.

[0731] In yet another embodiment, the invention concerns agonists andantagonists of the native PRO285 and PRO286 polypeptides. In aparticular embodiment, the agonist or antagonist is an anti-PRO285 oranti-PRO286 antibody.

[0732] In a further embodiment, the invention concerns screening assaysto identify agonists or antagonists of the native PRO285 and PRO286polypeptides.

[0733] In a still further embodiment, the invention concerns acomposition comprising a PRO285 or PRO286 polypeptide, or an agonist orantagonist as hereinabove defined, in combination with apharmaceutically acceptable carrier.

[0734] The invention further concerns a composition comprising anantibody specifically binding a PRO285 or PRO286 polypeptide, incombination with a pharmaceutically acceptable carrier.

[0735] The invention also concerns a method of treating septic shockcomprising administering to a patient an effective amount of anantagonist of a PRO285 or PRO286 polypeptide. In a specific embodiment,the antagonist is a blocking antibody specifically binding a nativePRO285 or PRO286 polypeptide.

[0736] 84. PRO213-1, PRO1330 and PRO1449

[0737] The present invention concerns compositions and methods for thediagnosis and treatment of neoplastic cell growth and proliferation inmammals, including humans. The present invention is based on theidentification of genes that are amplified in the genome of tumor cells.Such gene amplification is expected to be associated with theoverexpression of the gene product and contribute to tumorigenesis.Accordingly, the proteins encoded by the amplified genes are believed tobe useful targets for the diagnosis and/or treatment (includingprevention) of certain cancers, and may act as predictors of theprognosis of tumor treatment. In one embodiment, the present inventionprovides an isolated nucleic acid molecule comprising DNA encoding aPRO213-1, PRO1330 and/or PRO1449 polypeptide. In one aspect, theisolated nucleic acid comprises DNA encoding the PRO213-1, PRO1330and/or PRO1449 polypeptide having amino acid residues 1 to 295 of FIG.213 (SEQ ID NO:506), 20 to 273 of FIG. 215 (SEQ ID NO:508) and 20 to 273of FIG. 217 (SEQ ID NO:510), respectively, or is complementary to suchencoding nucleic acid sequence, and remains stably bound to it under atleast moderate, and optionally, under high stringency conditions. Theisolated nucleic acid sequence may comprise the cDNA insert of thevector designated as DNA30943-1163 (ATCC 209791) deposited on Apr. 21,1998; DNA64907-1163-1 (ATCC 203242) deposited on Sep. 9, 1998 and/orDNA64908-1163-1 (ATCC 203243) deposited on Sep. 9, 1998.

[0738] In another embodiment, the present invention comprises anisolated nucleic acid molecule having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding aPRO213-1, PRO1330 and/or PRO1449 polypeptide having amino acid residues1 to 295 of FIG. 213 (SEQ ID NO:506), 20 to 273 of FIG. 215 (SEQ IDNO:508) and 20 to 273 of FIG. 217 (SEQ ID NO:510), respectively; or (b)the complement of the DNA molecule of (a).

[0739] In another embodiment, the invention provides an isolatedPRO213-1, PRO1330 and/or PRO1449 polypeptide. In particular, theinvention provides isolated native sequence PRO213-1, PRO1330 and/orPRO1449 polypeptide, which in one embodiment, includes an amino acidsequence comprising residues 1 to 295 of FIG. 213 (SEQ ID NO:506), 20 to273 of FIG. 215 (SEQ ID NO:508) or 20 to 273 of FIG. 217 (SEQ IDNO:510), respectively. Optionally, the PRO213-1, PRO1330 and/or PRO1449polypeptide is obtained or obtainable by expressing the polypeptideencoded by the cDNA insert of the DNA30943-1163 (ATCC 209791),DNA64907-1163-1 (ATCC 203242) or DNA64908-1163-1 (ATCC 203243).

[0740] In another aspect, the invention provides an isolated PRO213-1,PRO1330, and/or PRO1449 polypeptide, comprising an amino acid sequencehaving at least about 80% sequence identity, preferably at least about85% sequence identity, more preferably at least about 95% sequenceidentity to amino acid residues 1 to 295 of FIG. 213 (SEQ ID NO:506), 20to 273 of FIG. 215 (SEQ ID NO:508) or 20 to 273 of FIG. 217 (SEQ IDNO:510), inclusive.

[0741] In yet another embodiment, the invention provides an isolatedPRO213-1, PRO1330, and/or PRO1449 polypeptide, comprising the amino acidresidues 1 to 295 of FIG. 213 (SEQ ID NO:506), 20 to 273 of FIG. 215(SEQ ID NO:508) or 20 to 273 of FIG. 217 (SEQ ID NO:510), or a fragmentthereof sufficient to provide a binding site for an anti-PRO213-1,anti-PRO1330and/or anti-PRO1449antibody. Preferably, the PRO213-1,PRO1330, and/or PRO1449 fragment retains a qualitative biologicalactivity of a native PRO213-1, PRO1330, and/or PRO1449 polypeptide.

[0742] In a further aspect, the invention concerns an isolated PRO213-1,PRO1330, and/or PRO1449 polypeptide, comprising an amino acid sequencescoring at least about 80% positives, preferably at least about 85%positives, more preferably at least about 90% positives, most preferablyat least about 95% positives when compared with the amino acid sequenceof residues 1 to 295 of FIG. 213 (SEQ ID NO:506), 20 to 273 of FIG. 215(SEQ ID NO:508) and 20 to 273 of FIG. 217 (SEQ ID NO:510), respectively.

[0743] In still a further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with: (a) a DNA molecule encoding a PRO213-1, PRO1330, and/orPRO1449 polypeptide having the amino acid residues from 1 to 295 of FIG.213 (SEQ ID NO:506), 20 to 273 of FIG. 215 (SEQ ID NO:508) and 20 to 273of FIG. 217 (SEQ ID NO:510), respectively; or the complement of the DNAmolecule of (a), and if said test DNA molecule has at least about an 80%sequence identity to (a) or (b), (ii) culturing a host cell comprisingsaid test DNA molecule under conditions suitable for the expression ofsaid polypeptide, and (iii) recovering said polypeptide from the cellculture.

[0744] In one embodiment, the present invention concerns an isolatedantibody which binds a PRO213-1, PRO1330 and/or PRO1449 polypeptide. Inone aspect, the antibody induces death of a cell overexpressing aPRO213-1, PRO1330 and/or PRO1449 polypeptide. In another aspect, theantibody is a monoclonal antibody, which preferably has nonhumancomplementarity determining region (CDR) residues and human frameworkregion (FR) residues. The antibody may be labeled and may be immobilizedon a solid support. In a further aspect, the antibody is an antibodyfragment, a single-chain antibody, or an anti-idiotypic antibody.

[0745] In another embodiment, the invention concerns a compositioncomprising an antibody which binds a PRO213-1, PRO1330 and/or PRO1449polypeptide in admixture with a pharmaceutically acceptable carrier. Inone aspect, the composition comprises a therapeutically effective amountof the antibody. In another aspect, the composition comprises a furtheractive ingredient, which may, for example, be a further antibody or acytotoxic or chemotherapeutic agent. Preferably, the composition issterile.

[0746] In a further embodiment, the invention concerns nucleic acidencoding an anti-PRO213-1, anti-PRO1330 and/or anti-PRO1449 antibody,and vectors and recombinant host cells comprising such nucleic acid.

[0747] The invention further concerns antagonists and agonists of aPRO213-1, PRO1330 and/or PRO1449 polypeptide that inhibit one or more ofthe functions or activities of the PRO213-1, PRO1330 and/or PRO1449polypeptide.

[0748] In a further embodiment, the invention concerns isolated nucleicacid molecules that hybridize to the complement of the nucleic acidmolecules encoding the PRO213-1, PRO1330 and/or PRO1449 polypeptides.The nucleic acid preferably is DNA, and hybridization preferably occursunder stringent conditions. Such nucleic acid molecules can act asantisense molecules of the amplified genes identified herein, which, inturn, can find use in the modulation of the respective amplified genes,or as antisense primers in amplification reactions. Furthermore, suchsequences can be used as part of ribozyme and/or triple helix sequencewhich, in turn, may be used in regulation of the amplified genes.

[0749] In another embodiment, the invention concerns a method fordetermining the presence of a PRO213-1, PRO1330 and/or PRO1449polypeptide comprising exposing a cell suspected of containing thePRO213-1, PRO1330 and/or PRO1449 polypeptide to an anti-PRO213-1,PRO1330 and/or PRO1449 antibody and determining binding of the antibodyto the cell.

[0750] In yet another embodiment, the present invention concerns amethod of diagnosing tumor in a mammal, comprising detecting the levelof expression of a gene encoding a PRO213-1, PRO1330 and/or PRO1449polypeptide (a) in a test sample of tissue cells obtained from themammal, and (b) in a control sample of known normal tissue cells of thesame cell type, wherein a higher expression level in the test sampleindicates the presence of tumor in the mammal from which the test tissuecells were obtained.

[0751] In another embodiment, the present invention concerns a method ofdiagnosing tumor in a mammal, comprising (a) contacting ananti-PRO213-1, anti-PRO1330 and/or anti-PRO1449 antibody with a testsample of tissue cells obtained from the mammal, and (b) detecting theformation of a complex between the anti- PRO213-1, anti-PRO1330 and/oranti-PRO1449 antibody and the PRO213-1, PRO1330 and/or PRO1449polypeptide in the test sample. The detection may be qualitative orquantitative, and may be performed in comparison with monitoring thecomplex formation in a control sample of known normal tissue cells ofthe same Gus cell type. A larger quantity of complexes formed in thetest sample indicates the presence of tumor in the mammal from which thetest tissue cells were obtained. The antibody preferably carries adetectable label. Complex formation can be monitored, for example, bylight microscopy, flow cytometry, fluorimetry, or other techniques knownin the art. The test sample is usually obtained from an individualsuspected to have neoplastic cell growth or proliferation (e.g.cancerous cells).

[0752] In another embodiment, the present invention concerns a cancerdiagnostic kit, comprising an anti-PRO213-1, anti-PRO1330 and/oranti-PRO1449 antibody and a carrier (e.g. a buffer) in suitablepackaging. The kit preferably contains instructions for using theantibody to detect the PRO213-1, PRO1330 and/or PRO1449 polypeptide.

[0753] In yet another embodiment, the invention concerns a method forinhibiting the growth of tumor cells comprising exposing a cell whichoverexpresses a PRO213-1, PRO1330 and/or PRO1449 polypeptide to aneffective amount of an agent inhibiting the expression and/or activityof the PRO213-1, PRO1330 and/or PRO1449 polypeptide. The agentpreferably is an anti-PRO213-1, anti-PRO1330 and/or anti-PRO1449antibody, a small organic and inorganic molecule, peptide,phosphopeptide, antisense or ribozyme molecule, or a triple helixmolecule. In a specific aspect, the agent, e.g. anti-PRO213-1,anti-PRO1330 and/or anti-PRO1449 antibody induces cell death. In afurther aspect, the tumor cells are further exposed to radiationtreatment and/or a cytotoxic or chemotherapeutic agent.

[0754] In a further embodiment, the invention concerns an article ofmanufacture, comprising:

[0755] a) a container;

[0756] b) a label on the container; and

[0757] c) a composition comprising an active agent contained within thecontainer; wherein the composition is effective for inhibiting thegrowth of tumor cells, the label on the container indicates that thecomposition can be used for treating conditions characterized byoverexpression of a PRO213-1, PRO1330 and/or PRO1449 polypeptide, andthe active agent in the composition is an agent inhibiting theexpression and/or activity of the PRO213-1,PRO1330and/orPRO1449polypeptide. In a preferred aspect, the active agentis an anti-PRO213-1, anti-PRO1330 and/or anti-PRO1449 antibody.

[0758] In yet a further embodiment, the invention provides a method foridentifying a compound capable of inhibiting the expression and/oractivity of a PRO213-1, PRO1330 and/or PRO1449 polypeptide, comprisingcontacting a candidate compound with a PRO213-1, PRO1330 and/or PRO1449polypeptide under conditions and for a time sufficient to allow thesetwo components to interact. In a specific aspect, either the candidatecompound or the PRO213-1, PRO1330 and/or PRO1449 polypeptide isimmobilized on a solid support. In another aspect, the non-immobilizedcomponent carries a detectable label.

[0759] 85. PRO298

[0760] Applicants have identified a cDNA clone that encodes a novelpolypeptide. The DNA is designated in the present application as“DNA39975-1210”, encoding a novel multi-transmembrane protein, referredto as “PRO298”.

[0761] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA having at least about 80%, preferably atleast about 85%, more preferably at least about 90%, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding PRO298,comprising the sequence of amino acids 1 to 364 of FIG. 219 (SEQ IDNO:515), or (b) the complement of the DNA molecule of (a). In oneaspect, the isolated nucleic acid comprises DNA encoding a PRO298polypeptide having amino acid residues 1 to 364 of FIG. 219 (SEQ IDNO:515), or is complementary to such encoding nucleic acid sequence, andremains stably bound to it under at least moderate, and optionally,under high stringency conditions.

[0762] In a further embodiment, the invention concerns an isolatednucleic acid molecule comprising DNA having at least an 80% sequenceidentity to (a) a DNA molecule encoding the same mature polypeptideencoded by the human protein cDNA in ATCC Deposit No. 209783(DNA39975-1210), or (b) the complement of the DNA molecule of (a).

[0763] In a still further embodiment, the invention concerns nucleicacid which comprises a DNA molecule encoding the same mature polypeptideencoded by the human protein cDNA in ATCC Deposit No. 209783(DNA39975-1210).

[0764] In another embodiment, the invention provides isolated PRO298polypeptide. In particular, the invention provides isolated nativesequence PRO298 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 364 of FIG. 219 (SEQ ID NO:515).

[0765] In another embodiment, the invention provides an expressedsequence tag (EST) designated DNA26832 comprising the nucleotidesequence of FIG. 220 (SEQ ID NO:516).

[0766] 86. PRO337

[0767] Applicants have identified a cDNA clone (DNA43316-1237) thatencodes a novel polypeptide, designated in the present application as“PRO337”.

[0768] In one embodiment, the invention provides an isolated nucleicacid molecule having at least about 80% sequence identity to (a) a DNAmolecule encoding a PRO337 polypeptide comprising the sequence of aminoacids 1 to 344 of FIG. 222 (SEQ ID NO:523), or (b) the complement of theDNA molecule of (a). The sequence identity preferably is about 85%, morepreferably about 90%, most preferably about 95%. In one aspect, theisolated nucleic acid has at least about 80%, preferably at least about85%, more preferably at least about 90%, and most preferably at leastabout 95 (including 96, 97, 98 and 99%) sequence identity with apolypeptide having amino acid residues 1 to 344 of FIG. 222 (SEQ IDNO:523). Preferably, the highest degree of sequence identity occurswithin the immunoglobulin and major histocompatibility domains (aminoacids 113 to 130 of FIG. 222, SEQ ID NO:523).

[0769] In a further embodiment, the isolated nucleic acid moleculecomprises DNA encoding a neurotrimin polypeptide having amino acidresidues 1 to 344 of FIG. 222 (SEQ ID NO:523), or is complementary tosuch encoding nucleic acid sequence, and remains stably bound to itunder at least moderate, and optionally, under high stringencyconditions. In another aspect, the invention provides a nucleic acid ofthe full length protein of clone DNA43316-1237, deposited with the ATCCunder accession number ATCC 209487, alternatively the coding sequence ofclone DNA43316-1237, deposited under accession number ATCC 209487.

[0770] In yet another embodiment, the invention provides isolated PRO337polypeptide. In particular, the invention provides isolated nativesequence PRO337 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 344 of FIG. 222 (SEQ ID NO:523).Native PRO337 polypeptides with or without the native signal sequence(amino acids 1 to about 28 in FIG. 222 (SEQ ID NO:523), and with orwithout the initiating methionine are specifically included.Alternatively, the invention provides a PRO337 polypeptide encoded bythe nucleic acid deposited under accession number ATCC 209487.

[0771] In yet another embodiment, the invention provides an expressedsequence tag (EST) comprising the nucleotide sequences identified inFIG. 223 as DNA42301 (SEQ ID NO:524).

[0772] 87. PRO403

[0773] Applicants have identified a cDNA clone (DNA55800-1263) thatencodes a novel polypeptide, designated in the present application as“PRO403”.

[0774] In one embodiment, the invention provides an isolated nucleicacid molecule having at least about 80% sequence identity to (a) a DNAmolecule encoding a PRO403 polypeptide comprising the sequence of aminoacids 1 to 736 of FIG. 225 (SEQ ID NO:526), or (b) the complement of theDNA molecule of (a). The sequence identity preferably is about 85%, morepreferably about 90%, most preferably about 95%. In one aspect, theisolated nucleic acid has at least about 80%, preferably at least about85%, more preferably at least about 90%, and most preferably at leastabout 95% sequence identity with a polypeptide having amino acidresidues 1 to 736 of FIG. 225 (SEQ ID NO:526). Preferably, the highestdegree of sequence identity occurs within: (1) the putativeN-glycosylatation sites (amino acid residues 132, 136, 177, 237, 282,349, 505, 598 and 606; (2) Cys residues conserved with the Kell bloodgroup protein family (amino acid residues 65, 70, 88 and 96) and theputative zinc binding motif (amino acid residues 570-579).

[0775] In a further embodiment, the isolated nucleic acid moleculecomprises DNA encoding a PRO403 polypeptide having amino acid residues 1to 736 of FIG. 225 (SEQ ID NO:526), or is complementary to such encodingnucleic acid sequence, and remains stably bound to it under at leastmoderate, and optionally, under high stringency conditions. In anotheraspect, the invention provides a nucleic acid of the full length proteinof clone DNA55800-1263, deposited with the ATCC under accession numberATCC 209680, alternatively the coding sequence of clone DNA55800-1263,deposited under accession number ATCC 209680.

[0776] In yet another embodiment, the invention provides isolated PRO403polypeptide. In particular, the invention provides isolated nativesequence PRO403 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 1 to 736 of FIG. 225 (SEQ ID NO:526).Native PRO403 polypeptides with or the initiating methionine arespecifically included. Alternatively, the invention provides a PRO403polypeptide encoded by the nucleic acid deposited under accession numberATCC 209680.

[0777] In yet another embodiment, the invention provides an expressedsequence tag (EST) and other sequence fragments comprising thenucleotide sequences identified herein as DNA34415 (FIG. 226; SEQ IDNO:527); DNA49830 (FIG. 227; SEQ ID NO:528) and DNA49831 (FIG. 228; SEQID NO:529).

[0778] 88. Additional Embodiments

[0779] In other embodiments of the present invention, the inventionprovides vectors comprising DNA encoding any of the herein describedpolypeptides. Host cell comprising any such vector are also provided. Byway of example, the host cells may be CHO cells, E. coli, or yeast. Aprocess for producing any of the herein described polypeptides isfurther provided and comprises culturing host cells under conditionssuitable for expression of the desired polypeptide and recovering thedesired polypeptide from the cell culture.

[0780] In other embodiments, the invention provides chimeric moleculescomprising any of the herein described polypeptides fused to aheterologous polypeptide or amino acid sequence. Example of suchchimeric molecules comprise any of the herein described polypeptidesfused to an epitope tag sequence or a Fc region of an immunoglobulin.

[0781] In another embodiment, the invention provides an antibody whichspecifically binds to any of the above or below described polypeptides.Optionally, the antibody is a monoclonal antibody, humanized antibody,antibody fragment or single-chain antibody.

[0782] In yet other embodiments, the invention provides oligonucleotideprobes useful for isolating genomic and cDNA nucleotide sequences or asantisense probes, wherein those probes may be derived from any of theabove or below described nucleotide sequences.

[0783] In other embodiments, the invention provides an isolated nucleicacid molecule comprising a nucleotide sequence that encodes a PROpolypeptide.

[0784] In one aspect, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% sequence identity,preferably at least about 81% sequence identity, more preferably atleast about 82% sequence identity, yet more preferably at least about83% sequence identity, yet more preferably at least about 84% sequenceidentity, yet more preferably at least about 85% sequence identity, yetmore preferably at least about 86% sequence identity, yet morepreferably at least about 87% sequence identity, yet more preferably atleast about 88% sequence identity, yet more preferably at least about89% sequence identity, yet more preferably at least about 90% sequenceidentity, yet more preferably at least about 91% sequence identity, yetmore preferably at least about 92% sequence identity, yet morepreferably at least about 93% sequence identity, yet more preferably atleast about 94% sequence identity, yet more preferably at least about95% sequence identity, yet more preferably at least about 96% sequenceidentity, yet more preferably at least about 97% sequence identity, yetmore preferably at least about 98% sequence identity and yet morepreferably at least about 99% sequence identity to (a) a DNA moleculeencoding a PRO polypeptide having a full-length amino acid sequence asdisclosed herein, an amino acid sequence lacking the signal peptide asdisclosed herein, an extracellular domain of a transmembrane protein,with or without the signal peptide, as disclosed herein or any otherspecifically defined fragment of the full-length amino acid sequence asdisclosed herein, or (b) the complement of the DNA molecule of (a).

[0785] In other aspects, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% sequence identity,preferably at least about 81% sequence identity, more preferably atleast about 82% sequence identity, yet more preferably at least about83% sequence identity, yet more preferably at least about 84% sequenceidentity, yet more preferably at least about 85% sequence identity, yetmore preferably at least about 86% sequence identity, yet morepreferably at least about 87% sequence identity, yet more preferably atleast about 88% sequence identity, yet more preferably at least about89% sequence identity, yet more preferably at least about 90% sequenceidentity, yet more preferably at least about 91% sequence identity, yetmore preferably at least about 92% sequence identity, yet morepreferably at least about 93% sequence identity, yet more preferably atleast about 94: % sequence identity, yet more preferably at least about95% sequence identity, yet more preferably at least about 96% sequenceidentity, yet more preferably at least about 97% sequence identity, yetmore preferably at least about 98% sequence identity and yet morepreferably at least about 99% sequence identity to (a) a DNA moleculecomprising the coding sequence of a full-length PRO polypeptide cDNA asdisclosed herein, the coding sequence of a PRO polypeptide lacking thesignal peptide as disclosed herein, the coding sequence of anextracellular domain of a transmembrane PRO polypeptide, with or withoutthe signal peptide, as disclosed herein or the coding sequence of anyother specifically defined fragment of the full-length amino acidsequence as disclosed herein, or (b) the complement of the DNA moleculeof (a).

[0786] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising a nucleotide sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to (a) a DNAmolecule that encodes the same mature polypeptide encoded by any of thehuman protein cDNAs deposited with the ATCC as disclosed herein, or (b)the complement of the DNA molecule of (a).

[0787] Another aspect the invention provides an isolated nucleic acidmolecule comprising a nucleotide sequence encoding a PRO polypeptidewhich is either transmembrane domain-deleted or transmembranedomain-inactivated, or is complementary to such encoding nucleotidesequence, wherein the transmembrane domain(s) of such polypeptide aredisclosed herein. Therefore, soluble extracellular domains of the hereindescribed PRO polypeptides are contemplated.

[0788] Another embodiment is directed to fragments of a PRO polypeptidecoding sequence, or the complement thereof, that may find use as, forexample, hybridization probes, for encoding fragments of a PROpolypeptide that may optionally encode a polypeptide comprising abinding site for an anti-PRO antibody or as antisense oligonucleotideprobes. Such nucleic acid fragments are usually at least about 20nucleotides in length, preferably at least about 30 nucleotides inlength, more preferably at least about 40 nucleotides in length, yetmore preferably at least about 50 nucleotides in length, yet morepreferably at least about 60 nucleotides in length, yet more preferablyat least about 70 nucleotides in length, yet more preferably at leastabout 80 nucleotides in length, yet more preferably at least about 90nucleotides in length, yet more preferably at least about 100nucleotidesin length, yet more preferably at least about 110 nucleotides in length,yet more preferably at least about 120 nucleotides in length, yet morepreferably at least about 130 nucleotides in length, yet more preferablyat least about 140 nucleotides in length, yet more preferably at leastabout 150 nucleotides in length, yet more preferably at least about 160nucleotides in length, yet more preferably at least about 170nucleotides 2o in length, yet more preferably at least about 180nucleotides in length, yet more preferably at least about 190nucleotides in length, yet more preferably at least about 200nucleotides in length, yet more preferably at least about 250nucleotides in length, yet more preferably at least about 300nucleotides in length, yet more preferably at least about 350nucleotides in length, yet more preferably at least about 400nucleotides in length, yet more preferably at least about 450nucleotides in length, yet more preferably at least about 500nucleotides in length, yet more preferably at least about 600nucleotides in length, yet more preferably at least about 700nucleotides in length, yet more preferably at least about 800nucleotides in length, yet more preferably at least about 900nucleotides in length and yet more preferably at least about 1000nucleotides in length, wherein in this context the term “about” meansthe referenced nucleotide sequence length plus or minus 10% of thatreferenced length. It is noted that novel fragments of a PRO polypeptidea coding nucleotide sequence may be determined in a routine manner byaligning the PRO polypeptide-encoding nucleotide sequence with otherknown nucleotide sequences using any of a number of well known sequencealignment programs and determining which PRO polypeptide-encodingnucleotide sequence fragment(s) are novel. All of such PROpolypeptide-encoding nucleotide sequences are contemplated herein. Alsocontemplated are the PRO polypeptide fragments encoded by thesenucleotide molecule fragments, preferably those PRO polypeptidefragments that comprise a binding site for an anti-PRO antibody.

[0789] In another embodiment, the invention provides isolated PROpolypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0790] In a certain aspect, the invention concerns an isolated PROpolypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity,. yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to a PROpolypeptide having a full-length amino acid sequence as disclosedherein, an amino acid sequence lacking the signal peptide as disclosedherein, an extracellular domain of a transmembrane protein, with orwithout the signal peptide, as disclosed herein or any otherspecifically defined fragment of the full-length amino acid sequence asdisclosed herein.

[0791] In a further aspect, the invention concerns an isolated PROpolypeptide comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to an aminoacid sequence encoded by any of the human protein cDNAs deposited withthe ATCC as disclosed herein.

[0792] In a further aspect, the invention concerns an isolated PROpolypeptide comprising an amino acid sequence scoring at least about 80%positives, preferably at least about 81% positives, more preferably atleast about 82% positives, yet more preferably at least about 83%positives, yet more preferably at least about 84% positives, yet morepreferably at least about 85% positives, yet more preferably at leastabout 86% positives, yet more preferably at least about 87% positives,yet more preferably at least about 88% positives, yet more preferably atleast about 89% positives, yet more preferably at least about 90%positives, yet more preferably at least about 91% positives, yet morepreferably at least about 92% positives, yet more preferably at leastabout 93% positives, yet more preferably at least about 94% positives,yet more preferably at least about 95% positives, yet more preferably atleast about 96% positives, yet more preferably at least about 97%positives, yet more preferably at least about 98% positives and yet morepreferably at least about 99% positives when compared with the aminoacid sequence of a PRO polypeptide having a full-length amino acidsequence as disclosed herein, an amino acid sequence lacking the signalpeptide as disclosed herein, an extracellular domain of a transmembraneprotein, with or without the signal peptide, as disclosed herein or anyother specifically defined fragment of the full-length amino acidsequence as disclosed herein.

[0793] In a specific aspect, the invention provides an isolated PROpolypeptide without the N-terminal signal sequence and/or the initiatingmethionine and is encoded by a nucleotide sequence that encodes such anamino acid sequence as hereinbefore described. Processes for producingthe same are also herein described, wherein those processes compriseculturing a host cell comprising a vector which comprises theappropriate encoding nucleic acid molecule under conditions suitable forexpression of the PRO polypeptide and recovering the PRO polypeptidefrom the cell culture.

[0794] Another aspect the invention provides an isolated PRO polypeptidewhich is either transmembrane domain-deleted or transmembranedomain-inactivated. Processes for producing the same are also hereindescribed, wherein those processes comprise culturing a host cellcomprising a vector which comprises the appropriate encoding nucleicacid molecule under conditions suitable for expression of the PROpolypeptide and recovering the PRO polypeptide from the cell culture.

[0795] In yet another embodiment, the invention concerns agonists andantagonists of a native PRO polypeptide as defined herein. In aparticular embodiment, the agonist or antagonist is an anti-PRO antibodyor a small molecule.

[0796] In a further embodiment, the invention concerns a method ofidentifying agonists or antagonists to a PRO polypeptide which comprisecontacting the PRO polypeptide with a candidate molecule and monitoringa biological activity mediated by said PRO polypeptide. Preferably, thePRO polypeptide is a native PRO polypeptide.

[0797] In a still further embodiment, the invention concerns acomposition of matter comprising a PRO polypeptide, or an agonist orantagonist of a PRO polypeptide as herein described, or an anti-PROantibody, in combination with a carrier. Optionally, the carrier is apharmaceutically acceptable carrier.

[0798] Another embodiment of the present invention is directed to theuse of a PRO polypeptide, or an agonist or antagonist thereof ashereinbefore described, or an anti-PRO antibody, for the preparation ofa medicament useful in the treatment of a condition which is responsiveto the PRO polypeptide, an agonist or antagonist thereof or an anti-PROantibody.

BRIEF DESCRIPTION OF THE DRAWINGS

[0799]FIG. 1 shows a nucleotide sequence (SEQ ID NO:1) of a nativesequence PRO213 cDNA, wherein SEQ ID NO:1 is a clone designated hereinas “UNQ187” and/or “DNA30943-1163”.

[0800]FIG. 2 shows the amino acid sequence (SEQ ID NO:2) derived fromthe coding sequence of SEQ ID NO:1 shown in FIG. 1.

[0801]FIG. 3 shows a nucleotide sequence (SEQ ID NO:6) of a nativesequence PRO274 cDNA, wherein SEQ ID NO:6 is a clone designated hereinas “UNQ241” and/or “DNA39987-1184”.

[0802]FIG. 4 shows the amino acid sequence (SEQ ID NO:7) derived fromthe coding sequence of SEQ ID NO:6 shown in FIG. 3.

[0803]FIG. 5 shows an EST nucleotide sequence designated herein asDNA17873 (SEQ ID NO:8).

[0804]FIG. 6 shows an EST nucleotide sequence designated herein asDNA36157 (SEQ ID NO:9).

[0805]FIG. 7 shows an EST nucleotide sequence designated herein asDNA28929 (SEQ ID NO:10).

[0806]FIG. 8 shows a nucleotide sequence (SEQ ID NO:18) of a nativesequence PRO300 cDNA, wherein SEQ ID NO:18 is a clone designated hereinas “UNQ263” and/or “DNA40625-1189”.

[0807]FIG. 9 shows the amino acid sequence (SEQ ID NO:19) derived fromthe coding sequence of SEQ ID NO:18 shown in FIG. 8.

[0808]FIG. 10 shows a nucleotide sequence (SEQ ID NO:27) of a nativesequence PRO284 cDNA, wherein SEQ ID NO:27 is a clone designated hereinas “UNQ247” and/or “DNA23318-1211”.

[0809]FIG. 11 shows the amino acid sequence (SEQ ID NO:28) derived fromthe coding sequence of SEQ ID NO:27 shown in FIG. 10.

[0810]FIG. 12 shows an EST nucleotide sequence designated herein asDNA12982 (SEQ ID NO:29).

[0811]FIG. 13 shows an EST nucleotide sequence designated herein asDNA15886 (SEQ ID NO:30).

[0812]FIG. 14 shows a nucleotide sequence (SEQ ID NO:35) of a nativesequence PRO296 cDNA, wherein SEQ ID NO:35 is a clone designated hereinas “UNQ260” and/or “DNA39979-1213”.

[0813]FIG. 15 shows the amino acid sequence (SEQ ID NO:36) derived fromthe coding sequence of SEQ ID NO:35 shown in FIG. 14.

[0814]FIG. 16 shows an EST nucleotide sequence designated herein asDNA3020 (SEQ ID NO:37).

[0815]FIG. 17 shows an EST nucleotide sequence designated herein asDNA21971 (SEQ ID NO:38).

[0816]FIG. 18 shows an EST nucleotide sequence designated herein asDNA29037 (SEQ ID NO:39).

[0817]FIG. 19 shows a nucleotide sequence (SEQ ID NO:44) of a nativesequence PRO329 cDNA, wherein 20SEQ ID NO:44 is a clone designatedherein as “UNQ291” and/or “DNA40594-1233”.

[0818]FIG. 20 shows the amino acid sequence (SEQ ID NO:45) derived fromthe coding sequence of SEQ ID NO:44 shown in FIG. 19.

[0819]FIG. 21 shows a nucleotide sequence (SEQ ID NO:51) of a nativesequence PRO362 cDNA, wherein SEQ ID NO:51 is a clone designated hereinas “UNQ317” and/or “DNA4541-1251”.

[0820]FIG. 22 shows the amino acid sequence (SEQ ID NO:52) derived fromthe coding sequence of SEQ ID NO:51 shown in FIG. 21.

[0821]FIG. 23 shows a nucleotide sequence (SEQ ID NO:58) of a nativesequence PRO363 cDNA, wherein SEQ ID NO:58 is a clone designated hereinas “UNQ318” and/or “DNA45419-1252”.

[0822]FIG. 24 shows the amino acid sequence (SEQ ID NO:59) derived fromthe coding sequence of SEQ ID NO:58 shown in FIG. 23.

[0823]FIG. 25 shows a nucleotide sequence (SEQ ID NO:63) of a nativesequence PRO868 cDNA, wherein SEQ ID NO:63 is a clone designated hereinas “UNQ437” and/or “DNA52594-1270”.

[0824]FIG. 26 shows the amino acid sequence (SEQ ID NO:64) derived fromthe coding sequence of SEQ ID NO:63 shown in FIG. 25.

[0825]FIG. 27 shows a nucleotide sequence (SEQ ID NO:68) of a nativesequence PRO382 cDNA, wherein SEQ ID NO:68 is a clone designated hereinas “UNQ323” and/or “DNA45234-1277”.

[0826]FIG. 28 shows the amino acid sequence (SEQ ID NO:69) derived fromthe coding sequence of SEQ ID NO:68 shown in FIG. 27.

[0827]FIG. 29 shows a nucleotide sequence (SEQ ID NO:73) of a nativesequence PRO545 cDNA, wherein SEQ ID NO:73 is a clone designated hereinas “UNQ346” and/or “DNA49624-1279”.

[0828]FIG. 30 shows the amino acid sequence (SEQ ID NO:74) derived fromthe coding sequence of SEQ ID NO:73 shown in FIG. 29.

[0829]FIG. 31 shows an EST nucleotide sequence designated herein asDNA13217 (SEQ ID NO:75).

[0830]FIG. 32 shows a nucleotide sequence (SEQ ID NO:84) of a nativesequence PRO617 cDNA, wherein SEQ ID NO:84 is a clone designated hereinas “UNQ353” and/or “DNA48309-1280”.

[0831]FIG. 33 shows the amino acid sequence (SEQ ID NO:85) derived fromthe coding sequence of SEQ ID NO:84 shown in FIG. 32.

[0832]FIG. 34 shows a nucleotide sequence (SEQ ID NO:89) of a nativesequence PRO700 cDNA, wherein SEQ ID NO:89 is a clone designated hereinas “UNQ364” and/or “DNA46776-1284”.

[0833]FIG. 35 shows the amino acid sequence (SEQ ID NO:90) derived fromthe coding sequence of SEQ ID NO:89 shown in FIG. 34.

[0834]FIG. 36 shows a nucleotide sequence (SEQ ID NO:96) of a nativesequence PRO702 cDNA, wherein SEQ ID NO:96 is a clone designated hereinas “UNQ366” and/or “DNA50980-1286”.

[0835]FIG. 37 shows the amino acid sequence (SEQ ID NO:97) derived fromthe coding sequence of SEQ ID NO:96 shown in FIG. 36.

[0836]FIG. 38 shows a nucleotide sequence (SEQ ID NO:101) of a nativesequence PRO703 cDNA, wherein SEQ ID NO:101 is a clone designated hereinas “UNQ367” and/or “DNA50913-1287”.

[0837]FIG. 39 shows the amino acid sequence (SEQ ID NO:102) derived fromthe coding sequence of SEQ ID NO:101 shown in FIG. 38.

[0838]FIG. 40 shows a nucleotide sequence (SEQ ID NO:108) of a nativesequence PRO705 cDNA, wherein SEQ ID NO:108 is a clone designated hereinas “UNQ369” and/or “DNA50914-1289”.

[0839]FIG. 41 shows the amino acid sequence (SEQ ID NO:109) derived fromthe coding sequence of SEQ ID NO:108 shown in FIG. 40.

[0840] FIGS. 42A-B show a nucleotide sequence (SEQ ID NO:113) of anative sequence PRO708 cDNA, wherein SEQ ID NO:113 is a clone designatedherein as “UNQ372” and/or “DNA48296-1292”.

[0841]FIG. 43 shows the amino acid sequence (SEQ ID NO:114) derived fromthe coding sequence of SEQ ID NO:113 shown in FIGS. 42A-B.

[0842]FIG. 44 shows a nucleotide sequence (SEQ ID NO:118) of a nativesequence PRO320 cDNA, wherein SEQ ID NO:118 is a clone designated hereinas “UNQ281” and/or “DNA32284-1307”.

[0843]FIG. 45 shows the amino acid sequence (SEQ ID NO:119) derived fromthe coding sequence of SEQ ID NO:118 shown in FIG. 44.

[0844]FIG. 46 shows a nucleotide sequence (SEQ ID NO:123) of a nativesequence PRO324 cDNA, wherein SEQ ID NO:123 is a clone designated hereinas “UNQ285” and/or “DNA36343-1310”.

[0845]FIG. 47 shows the amino acid sequence (SEQ ID NO:124) derived fromthe coding sequence of SEQ ID NO:123 shown in FIG. 46.

[0846]FIG. 48 shows a nucleotide sequence (SEQ ID NO:131) of a nativesequence PRO351 cDNA, wherein SEQ ID NO:131 is a clone designated hereinas “UNQ308” and/or “DNA40571-1315”.

[0847]FIG. 49 shows the amino acid sequence (SEQ ID NO:132) derived fromthe coding sequence of SEQ ID NO:131 shown in FIG. 48.

[0848]FIG. 50 shows a nucleotide sequence (SEQ ID NO:136) of a nativesequence PRO352 cDNA, wherein SEQ ID NO:136 is a clone designated hereinas “UNQ309” and/or “DNA41386-1316”.

[0849]FIG. 51 shows the amino acid sequence (SEQ ID NO:137) derived fromthe coding sequence of SEQ ID NO:136 shown in FIG. 50.

[0850]FIG. 52 shows a nucleotide sequence (SEQ ID NO:144) of a nativesequence PRO381 cDNA, wherein SEQ ID NO:144 is a clone designated hereinas “UNQ322” and/or “DNA44194-1317”.

[0851]FIG. 53 shows the amino acid sequence (SEQ ID NO:145) derived fromthe coding sequence of SEQ ID NO:144 shown in FIG. 52.

[0852]FIG. 54 shows a nucleotide sequence (SEQ ID NO:149) of a nativesequence PRO386 cDNA, wherein SEQ ID NO:149 is a clone designated hereinas “UNQ326” and/or “DNA45415-1318”.

[0853]FIG. 55 shows the amino acid sequence (SEQ ID NO:150) derived fromthe coding sequence of SEQ ID NO:149 shown in FIG. 54.

[0854]FIG. 56 shows an EST nucleotide sequence designated herein asDNA23350 (SEQ ID NO:151).

[0855]FIG. 57 shows an EST nucleotide sequence designated herein asDNA23536 (SEQ ID NO:152).

[0856]FIG. 58 shows a nucleotide sequence (SEQ ID NO:156) of a nativesequence PRO540 cDNA, wherein SEQ ID NO:156 is a clone designated hereinas “UNQ341” and/or “DNA44189-1322”.

[0857]FIG. 59 shows the amino acid sequence (SEQ ID NO:157) derived fromthe coding sequence of SEQ ID NO:156 shown in FIG. 58.

[0858]FIG. 60 shows a nucleotide sequence (SEQ ID NO:161) of a nativesequence PRO615 cDNA, wherein SEQ ID NO:161 is a clone designated hereinas “UNQ352” and/or “DNA48304-1323”.

[0859]FIG. 61 shows the amino acid sequence (SEQ ID NO:162) derived fromthe coding sequence of SEQ ID NO:161 shown in FIG. 60.

[0860]FIG. 62 shows a nucleotide sequence (SEQ ID NO:168) of a nativesequence PRO618 cDNA, wherein SEQ ID NO:168 is a clone designated hereinas “UNQ354” and/or “DNA49152-1324”.

[0861]FIG. 63 shows the amino acid sequence (SEQ ID NO:169) derived fromthe coding sequence of SEQ ID NO:168 shown in FIG. 62.

[0862]FIG. 64 shows an EST nucleotide sequence designated herein asDNA35597 (SEQ ID NO:170).

[0863]FIG. 65 shows a nucleotide sequence (SEQ ID NO:177) of a nativesequence PRO719 cDNA, wherein SEQ ID NO:177 is a clone designated hereinas “UNQ387” and/or “DNA49646-1327”.

[0864]FIG. 66 shows the amino acid sequence (SEQ ID NO:178) derived fromthe coding sequence of SEQ ID NO:177 shown in FIG. 65.

[0865]FIG. 67 shows a nucleotide sequence (SEQ ID NO:182) of a nativesequence PRO724 cDNA, wherein SEQ ID NO:182 is a clone designated hereinas “UNQ389” and/or “DNA49631-1328”.

[0866]FIG. 68 shows the amino acid sequence (SEQ ID NO:183) derived fromthe coding sequence of SEQ ID NO:182 shown in FIG. 67.

[0867]FIG. 69 shows a nucleotide sequence (SEQ ID NO:189) of a nativesequence PRO772 cDNA, wherein SEQ ID NO:189 is a clone designated hereinas “UNQ410” and/or “DNA49645-1347”.

[0868]FIG. 70 shows the amino acid sequence (SEQ ID NO:190) derived fromthe coding sequence of SEQ ID NO:189 shown in FIG. 69.

[0869]FIG. 71 shows an EST nucleotide sequence designated herein asDNA43509 (SEQ ID NO:191).

[0870]FIG. 72 shows a nucleotide sequence (SEQ ID NO:195) of a nativesequence PRO852 cDNA, wherein SEQ ID NO:195 is a clone designated hereinas “UNQ418” and/or “DNA45493-1349”.

[0871]FIG. 73 shows the amino acid sequence (SEQ ID NO:196) derived fromthe coding sequence of SEQ ID NO:195 shown in FIG. 72.

[0872]FIG. 74 shows a nucleotide sequence (SEQ ID NO:205) of a nativesequence PRO853 cDNA, wherein SEQ ID NO:205 is a clone designated hereinas “UNQ419” and/or “DNA48227-1350”.

[0873]FIG. 75 shows the amino acid sequence (SEQ ID NO:206) derived fromthe coding sequence of SEQ ID NO:205 shown in FIG. 74.

[0874]FIG. 76 shows a nucleotide sequence (SEQ ID NO:210) of a nativesequence PRO860 cDNA, wherein SEQ ID NO:210 is a clone designated hereinas “UNQ421” and/or “DNA41404-1352”.

[0875]FIG. 77 shows the amino acid sequence (SEQ ID NO:211) derived fromthe coding sequence of SEQ ID NO:210 shown in FIG. 76.

[0876]FIG. 78 shows a nucleotide sequence (SEQ ID NO:215) of a nativesequence PRO846 cDNA, wherein SEQ ID NO:215 is a clone designated hereinas “UNQ422” and/or “DNA44196-1353”.

[0877]FIG. 79 shows the amino acid sequence (SEQ ID NO:216) derived fromthe coding sequence of SEQ ID NO:215 shown in FIG. 78.

[0878]FIG. 80 shows a nucleotide sequence (SEQ ID NO:220) of a nativesequence PRO862 cDNA, wherein SEQ ID NO:220 is a clone designated hereinas “UNQ424” and/or “DNA52187-1354”.

[0879]FIG. 81 shows the amino acid sequence (SEQ ID NO:221) derived fromthe coding sequence of SEQ ID NO:220 shown in FIG. 80.

[0880]FIG. 82 shows a nucleotide sequence (SEQ ID NO:225) of a nativesequence PRO864 cDNA, wherein SEQ ID NO:225 is a clone designated hereinas “UNQ426” and/or “DNA48328-1355”.

[0881]FIG. 83 shows the amino acid sequence (SEQ ID NO:226) derived fromthe coding sequence of SEQ ID NO:225 shown in FIG. 82.

[0882]FIG. 84 shows a nucleotide sequence (SEQ ID NO:230) of a nativesequence PRO792 cDNA, wherein SEQ ID NO:230 is a clone designated hereinas “UNQ431” and/or “DNA56352-1358”.

[0883]FIG. 85 shows the amino acid sequence (SEQ ID NO:231) derived fromthe coding sequence of SEQ ID NO:230 shown in FIG. 84.

[0884]FIG. 86 shows a nucleotide sequence (SEQ ID NO:235) of a nativesequence PRO866 cDNA, wherein SEQ ID NO:235 is a clone designated hereinas “UNQ435” and/or “DNA53971-1359”.

[0885]FIG. 87 shows the amino acid sequence (SEQ ID NO:236) derived fromthe coding sequence of SEQ ID NO:235 shown in FIG. 86.

[0886]FIG. 88 shows a nucleotide sequence (SEQ ID NO:244) of a nativesequence PRO871 cDNA, wherein SEQ ID NO:244 is a clone designated hereinas “UNQ438” and/or “DNA50919-1361 ”.

[0887]FIG. 89 shows the amino acid sequence (SEQ ID NO:245) derived fromthe coding sequence of SEQ ID NO:244 shown in FIG. 88.

[0888]FIG. 90 shows a nucleotide sequence (SEQ ID NO:253) of a nativesequence PRO873 cDNA, wherein SEQ ID NO:253 is a clone designated hereinas “UNQ440” and/or “DNA44179-1362”.

[0889]FIG. 91 shows the amino acid sequence (SEQ ID NO:254) derived fromthe coding sequence of SEQ ID NO:253 shown in FIG. 90.

[0890]FIG. 92 shows a nucleotide sequence (SEQ ID NO:258) of a nativesequence PRO940 cDNA, wherein SEQ ID NO:258 is a clone designated hereinas “UNQ477” and/or “DNA54002-1367”.

[0891]FIG. 93 shows the amino acid sequence (SEQ ID NO:259) derived fromthe coding sequence of SEQ ID NO:258 shown in FIG. 92.

[0892]FIG. 94 shows a nucleotide sequence (SEQ ID NO:263) of a nativesequence PRO941 cDNA, wherein SEQ ID NO:263 is a clone designated hereinas “UNQ478” and/or “DNA53906-1368”.

[0893]FIG. 95 shows the amino acid sequence (SEQ ID NO:264) derived fromthe coding sequence of SEQ ID NO:263 shown in FIG. 94.

[0894]FIG. 96 shows an EST nucleotide sequence designated herein asDNA6415 (SEQ ID NO:265).

[0895]FIG. 97 shows a nucleotide sequence (SEQ ID NO:269) of a nativesequence PRO944 cDNA, wherein SEQ ID NO:269 is a clone designated hereinas “UNQ481” and/or “DNA52185-1370”.

[0896]FIG. 98 shows the amino acid sequence (SEQ ID NO:270) derived fromthe coding sequence of SEQ ID NO:269 shown in FIG. 97.

[0897]FIG. 99 shows an EST nucleotide sequence designated herein asDNA14007 (SEQ ID NO:271).

[0898]FIG. 100 shows an EST nucleotide sequence designated herein asDNA12773 (SEQ ID NO:272).

[0899]FIG. 101 shows an EST nucleotide sequence designated herein asDNA12746 (SEQ ID NO:273).

[0900]FIG. 102 shows an EST nucleotide sequence designated herein asDNA12834 (SEQ ID NO:274).

[0901]FIG. 103 shows an EST nucleotide sequence designated herein asDNA12846 (SEQ ID NO:275).

[0902]FIG. 104 shows an EST nucleotide sequence designated herein asDNA13104 (SEQ ID NO:276).

[0903]FIG. 105 shows an EST nucleotide sequence designated herein asDNA13259 (SEQ ID NO:277).

[0904]FIG. 106 shows an EST nucleotide sequence designated herein asDNA13959 (SEQ ID NO:278).

[0905]FIG. 107 shows an EST nucleotide sequence designated herein asDNA13961 (SEQ ID NO:279).

[0906]FIG. 108 shows a nucleotide sequence (SEQ ID NO:283) of a nativesequence PRO983 cDNA, wherein SEQ ID NO:283 is a clone designated hereinas “UNQ484” and/or “DNA53977-1371”.

[0907]FIG. 109 shows the amino acid sequence (SEQ ID NO:284) derivedfrom the coding sequence of SEQ ID NO:283 shown in FIG. 108.

[0908]FIG. 110 shows an EST nucleotide sequence designated herein asDNA17130 (SEQ ID NO:285).

[0909]FIG. 111 shows an EST nucleotide sequence designated herein asDNA23466 (SEQ ID NO:286).

[0910]FIG. 112 shows an EST nucleotide sequence designated herein asDNA26818 (SEQ ID NO:287).

[0911]FIG. 113 shows an EST nucleotide sequence designated herein asDNA37618 (SEQ ID NO:288).

[0912]FIG. 114 shows an EST nucleotide sequence designated herein asDNA41732 (SEQ ID NO:289).

[0913]FIG. 115 shows an EST nucleotide sequence designated herein asDNA45980 (SEQ ID NO:290).

[0914]FIG. 116 shows an EST nucleotide sequence designated herein asDNA46372 (SEQ ID NO:291).

[0915]FIG. 117 shows a nucleotide sequence (SEQ ID NO:295) of a nativesequence PRO1057 cDNA, wherein SEQ ID NO:295 is a clone designatedherein as “UNQ522” and/or “DNA57253-1382”.

[0916]FIG. 118 shows the amino acid sequence (SEQ ID NO:296) derivedfrom the coding sequence of SEQ ID NO:295 shown in FIG. 117.

[0917]FIG. 119 shows a nucleotide sequence (SEQ ID NO:300) of a nativesequence PRO1071 cDNA, wherein SEQ ID NO:300 is a clone designatedherein as “UNQ528” and/or “DNA58847-1383”.

[0918]FIG. 120 shows the amino acid sequence (SEQ ID NO:301) derivedfrom the coding sequence of SEQ ID NO:300 shown in FIG. 119.

[0919]FIG. 121 shows a nucleotide sequence (SEQ ID NO:302) of a nativesequence PRO1072 cDNA, wherein SEQ ID NO:302 is a clone designatedherein as “UNQ529” and/or “DNA58747-1384”.

[0920]FIG. 122 shows the amino acid sequence (SEQ ID NO:303) derivedfrom the coding sequence of SEQ ID NO:302 shown in FIG. 121.

[0921]FIG. 123 shows an EST nucleotide sequence designated herein asDNA40210 (SEQ ID NO:304).

[0922]FIG. 124 shows a nucleotide sequence (SEQ ID NO:308) of a nativesequence PRO1075 cDNA, wherein SEQ ID NO:308 is a clone designatedherein as “UNQ532” and/or “DNA57689-1385”.

[0923]FIG. 125 shows the amino acid sequence (SEQ ID NO:309) derivedfrom the coding sequence of SEQ ID NO:308 shown in FIG. 124.

[0924]FIG. 126 shows an EST nucleotide sequence designated herein asDNA13059 (SEQ ID NO:310).

[0925]FIG. 127 shows an EST nucleotide sequence designated herein asDNA19463 (SEQ ID NO:310).

[0926]FIG. 128 shows a nucleotide sequence (SEQ ID NO:321) of a nativesequence PRO181 cDNA, wherein SEQ ID NO:321 is a clone designated hereinas “UNQ155” and/or “DNA23330-1390”.

[0927]FIG. 129 shows the amino acid sequence (SEQ ID NO:322) derivedfrom the coding sequence of SEQ ID NO:321 shown in FIG. 128.

[0928]FIG. 130 shows an EST nucleotide sequence designated herein asDNA13242 (SEQ ID NO:323).

[0929]FIG. 131 shows a nucleotide sequence (SEQ ID NO:329) of a nativesequence PRO195 cDNA, wherein SEQ ID NO:329 is a clone designated hereinas “UNQ169” and/or “DNA26847-1395”.

[0930]FIG. 132 shows the amino acid sequence (SEQ ID NO:330) derivedfrom the coding sequence of SEQ ID NO:329 shown in FIG. 131.

[0931]FIG. 133 shows an EST nucleotide sequence designated herein asDNA15062 (SEQ ID NO:331).

[0932]FIG. 134 shows an EST nucleotide sequence designated herein asDNA13199 (SEQ ID NO:332).

[0933]FIG. 135 shows a nucleotide sequence (SEQ ID NO:336) of a nativesequence PRO865 cDNA, wherein SEQ ID NO:336 is a clone designated hereinas “UNQ434” and/or “DNA53974-1401”.

[0934]FIG. 136 shows the amino acid sequence (SEQ ID NO:337) derivedfrom the coding sequence of SEQ ID NO:336 shown in FIG. 135.

[0935]FIG. 137 shows an EST nucleotide sequence designated herein asDNA37642 (SEQ ID NO:338).

[0936]FIG. 138 shows a nucleotide sequence (SEQ ID NO:345) of a nativesequence PRO827 cDNA, wherein SEQ ID NO:345 is a clone designated hereinas “UNQ468” and/or “DNA57039-1402”.

[0937]FIG. 139 shows the amino acid sequence (SEQ ID NO:346) derivedfrom the coding sequence of SEQ ID NO:345 shown in FIG. 138.

[0938]FIG. 140 shows an EST nucleotide sequence designated herein asDNA47751 (SEQ ID NO:347).

[0939]FIG. 141 shows a nucleotide sequence (SEQ ID NO:351) of a nativesequence PRO114 cDNA, wherein SEQ ID NO:351 is a clone designated hereinas “UNQ557” and/or “DNA57033-1403”.

[0940]FIG. 142 shows the amino acid sequence (SEQ ID NO:352) derivedfrom the coding sequence of SEQ ID NO:351 shown in FIG. 141.

[0941]FIG. 143 shows an EST nucleotide sequence designated herein asDNA48466 (SEQ ID NO:353).

[0942]FIG. 144 shows a nucleotide sequence (SEQ ID NO:357) of a nativesequence PRO237 cDNA, wherein SEQ ID NO:357 is a clone designated hereinas “UNQ211 and/or “DNA34353-1428”.

[0943]FIG. 145 shows the amino acid sequence (SEQ ID NO:358) derivedfrom the coding sequence of SEQ ID NO:357 shown in FIG. 144.

[0944]FIG. 146 shows a nucleotide sequence (SEQ ID NO:362) of a nativesequence PRO541 cDNA, wherein SEQ ID NO:362 is a clone designated hereinas “UNQ342” and/or “DNA45417-1432”.

[0945]FIG. 147 shows the amino acid sequence (SEQ ID NO:363) derivedfrom the coding sequence of SEQ ID NO:362 shown in FIG. 146.

[0946]FIG. 148 shows a nucleotide sequence (SEQ ID NO:369) of a nativesequence PRO273 cDNA, wherein SEQ ID NO:369 is a clone designated hereinas “UNQ240” and/or “DNA39523-1192”.

[0947]FIG. 149 shows the amino acid sequence (SEQ ID NO:370) derivedfromthe coding sequence of SEQ ID NO:369 shown in FIG. 148.

[0948]FIG. 150 shows a nucleotide sequence (SEQ ID NO:374) of a nativesequence PRO701 cDNA, wherein SEQ ID NO:374 is a clone designated hereinas “UNQ365” and/or “DNA44205-1285”.

[0949]FIG. 151 shows the amino acid sequence (SEQ ID NO:375) derivedfrom the coding sequence of SEQ ID NO:374 shown in FIG. 150.

[0950]FIG. 152 shows a nucleotide sequence (SEQ ID NO:379) of a nativesequence PRO704 cDNA, wherein SEQ ID NO:379 is a clone designated hereinas “UNQ368” and/or “DNA50911-1288”.

[0951]FIG. 153 shows the amino acid sequence (SEQ ID NO:380) derivedfrom the coding sequence of SEQ ID NO:379 shown in FIG. 152.

[0952]FIG. 154 shows a nucleotide sequence (SEQ ID NO:384) of a nativesequence PRO706 cDNA, wherein SEQ ID NO:384 is a clone designated hereinas “UNQ370” and/or “DNA48329-1290”.

[0953]FIG. 155 shows the amino acid sequence (SEQ ID NO:385) derivedfrom the coding sequence of SEQ ID NO:384 shown in FIG. 154.

[0954]FIG. 156 shows a nucleotide sequence (SEQ ID NO:389) of a nativesequence PRO707 cDNA, wherein SEQ ID NO:389 is a clone designated hereinas “UNQ371” and/or “DNA48306-1291”.

[0955]FIG. 157 shows the amino acid sequence (SEQ ID NO:390) derivedfrom the coding sequence of SEQ ID NO:389 shown in FIG. 156.

[0956]FIG. 158 shows a nucleotide sequence (SEQ ID NO:394) of a nativesequence PRO322 cDNA, wherein SEQ ID NO:394 is a clone designated hereinas “UNQ283” and/or “DNA48336-1309”.

[0957]FIG. 159 shows the amino acid sequence (SEQ ID NO:395) derivedfrom the coding sequence of SEQ ID NO:394 shown in FIG. 158.

[0958]FIG. 160 shows a nucleotide sequence (SEQ ID NO:399) of a nativesequence PRO526 cDNA, wherein SEQ ID NO:399 is a clone designated hereinas “UNQ330” and/or “DNA44184-1319”.

[0959]FIG. 161 shows the amino acid sequence (SEQ ID NO:400) derivedfrom the coding sequence of SEQ ID NO:399 shown in FIG. 160.

[0960]FIG. 162 shows a nucleotide sequence (SEQ ID NO:404) of a nativesequence PRO531 cDNA, wherein SEQ ID NO:404 is a clone designated hereinas “UNQ332” and/or “DNA48314-1320”.

[0961]FIG. 163 shows the amino acid sequence (SEQ ID NO:405) derivedfrom the coding sequence of SEQ ID NO:404 shown in FIG. 162.

[0962]FIG. 164 shows a nucleotide sequence (SEQ ID NO:409) of a nativesequence PRO534 cDNA, wherein SEQ ID NO:409 is a clone designated hereinas “UNQ335” and/or “DNA48333-1321”.

[0963]FIG. 165 shows the amino acid sequence (SEQ ID NO:410) derivedfrom the coding sequence of SEQ ID NO:409 shown in FIG. 164.

[0964]FIG. 166 shows a nucleotide sequence (SEQ ID NO:414) of a nativesequence PRO697 cDNA, wherein SEQ ID NO:414 is a clone designated hereinas “UNQ361” and/or “DNA50920-1325”.

[0965]FIG. 167 shows the amino acid sequence (SEQ ID NO:415) derivedfrom the coding sequence of SEQ ID NO:414 shown in FIG. 166.

[0966]FIG. 168 shows a nucleotide sequence (SEQ ID NO:419) of a nativesequence PRO717 cDNA, wherein SEQ ID NO:419 is a clone designated hereinas “UNQ385” and/or “DNA50988-1326”.

[0967]FIG. 169 shows the amino acid sequence (SEQ ID NO:420) derivedfrom the coding sequence of SEQ ID NO:419 shown in FIG. 168.

[0968]FIG. 170 shows a nucleotide sequence (SEQ ID NO:424) of a nativesequence PRO731 cDNA, wherein SEQ ID NO:424 is a clone designated hereinas “UNQ395” and/or “DNA48331-1329”.

[0969]FIG. 171 shows the amino acid sequence (SEQ ID NO:425) derivedfrom the coding sequence of SEQ ID NO:424 shown in FIG. 170.

[0970]FIG. 172 shows a nucleotide sequence (SEQ ID NO:429) of a nativesequence PRO218 cDNA, wherein SEQ ID NO:429 is a clone designated hereinas “UNQ192” and/or “DNA30867-1335”.

[0971]FIG. 173 shows the amino acid sequence (SEQ ID NO:430) derivedfrom the coding sequence of SEQ ID NO:429 shown in FIG. 172.

[0972]FIG. 174 shows an EST nucleotide sequence designated herein asDNA14472 (SEQ ID NO:431).

[0973]FIG. 175 shows an EST nucleotide sequence designated herein asDNA15846 (SEQ ID NO:432).

[0974]FIG. 176 shows a nucleotide sequence (SEQ ID NO:436) of a nativesequence PRO768 cDNA, wherein SEQ ID NO:436 is a clone designated hereinas “UNQ406” and/or “DNA55737-1345”.

[0975]FIG. 177 shows the amino acid sequence (SEQ ID NO:437) derivedfrom the coding sequence of SEQ ID NO:436 shown in FIG. 176.

[0976]FIG. 178 shows a nucleotide sequence (SEQ ID NO:441) of a nativesequence PRO771 cDNA, wherein SEQ ID NO:441 is a clone designated hereinas “UNQ409” and/or “DNA49829-1346”.

[0977]FIG. 179 shows the amino acid sequence (SEQ ID NO:442) derivedfrom the coding sequence of SEQ ID NO:441 shown in FIG. 178.

[0978]FIG. 180 shows a nucleotide sequence (SEQ ID NO:446) of a nativesequence PRO733 cDNA, wherein SEQ ID NO:446 is a clone designated hereinas “UNQ411” and/or “DNA52196-1348”.

[0979]FIG. 181 shows the amino acid sequence (SEQ ID NO:447) derivedfrom the coding sequence of SEQ ID NO:446 shown in FIG. 180.

[0980]FIG. 182 shows a nucleotide sequence (SEQ ID NO:451) of a nativesequence PRO162 cDNA, wherein SEQ ID NO:451 is a clone designated hereinas “UNQ429” and/or “DNA56965-1356”.

[0981]FIG. 183 shows the amino acid sequence (SEQ ID NO:452) derivedfrom the coding sequence of SEQ ID NO:451 shown in FIG. 182.

[0982]FIG. 184 shows a nucleotide sequence (SEQ ID NO:453) of a nativesequence PRO788 cDNA, wherein SEQ ID NO:453 is a clone designated hereinas “UNQ430” and/or “DNA56405-1357”.

[0983]FIG. 185 shows the amino acid sequence (SEQ ID NO:454) derivedfrom the coding sequence of SEQ ID NO:453 shown in FIG. 184.

[0984]FIG. 186 shows a nucleotide sequence (SEQ ID NO:455) of a nativesequence PRO1008 cDNA, wherein SEQ ID NO:455 is a clone designatedherein as “UNQ492” and/or “DNA57530-1375”.

[0985]FIG. 187 shows the amino acid sequence (SEQ ID NO:456) derivedfrom the coding sequence of SEQ ID NO:455 shown in FIG. 186.

[0986]FIG. 188 shows an EST nucleotide sequence designated herein asDNA16508 (SEQ ID NO:457).

[0987]FIG. 189 shows a nucleotide sequence (SEQ ID NO:458) of a nativesequence PRO1012 cDNA, wherein SEQ ID NO:458 is a clone designatedherein as “UNQ495” and/or “DNA56439-1376”.

[0988]FIG. 190 shows the amino acid sequence (SEQ ID NO:459) derivedfrom the coding sequence of SEQ ID NO:458 shown in FIG. 189.

[0989]FIG. 191 shows a nucleotide sequence (SEQ ID NO:463) of a nativesequence PRO1014 cDNA, wherein SEQ ID NO:463 is a clone designatedherein as “UNQ497” and/or “DNA56409-1377”.

[0990]FIG. 192 shows the amino acid sequence (SEQ ID NO:464) derivedfrom the coding sequence of SEQ ID NO:463 shown in FIG. 191.

[0991]FIG. 193 shows a nucleotide sequence (SEQ ID NO:465) of a nativesequence PRO1017 cDNA, wherein SEQ ID NO:465 is a clone designatedherein as “UNQ500” and/or “DNA56112-1379”.

[0992]FIG. 194 shows the amino acid sequence (SEQ ID NO:466) derivedfrom the coding sequence of SEQ ID NO:465 shown in FIG. 193.

[0993]FIG. 195 shows a nucleotide sequence (SEQ ID NO:467) of a nativesequence PRO474 cDNA, wherein SEQ ID NO:467 is a clone designated hereinas “UNQ502” and/or “DNA56045-1380”.

[0994]FIG. 196 shows the amino acid sequence (SEQ ID NO:468) derivedfrom the coding sequence of SEQ ID NO:467 shown in FIG. 195.

[0995]FIG. 197 shows a nucleotide sequence (SEQ ID NO:469) of a nativesequence PRO1031 cDNA, wherein SEQ ID NO:469 is a clone designatedherein as “UNQ516” and/or “DNA59294-1381”.

[0996]FIG. 198 shows the amino acid sequence (SEQ ID NO:470) derivedfrom the coding sequence of SEQ ID NO:469 shown in FIG. 197.

[0997]FIG. 199 shows a nucleotide sequence (SEQ ID NO:471) of a nativesequence PRO938 cDNA, wherein SEQ ID NO:471 is a clone designated hereinas “UNQ475” and/or “DNA56433-1406”.

[0998]FIG. 200 shows the amino acid sequence (SEQ ID NO:472) derivedfrom the coding sequence of SEQ ID NO:471 shown in FIG. 199.

[0999]FIG. 201 shows a nucleotide sequence (SEQ ID NO:476) of a nativesequence PRO1082 cDNA, wherein SEQ ID NO:476 is a clone designatedherein as “UNQ539” and/or “DNA53912-1457”.

[1000]FIG. 202 shows the amino acid sequence (SEQ ID NO:477) derivedfrom the coding sequence of SEQ ID NO:476 shown in FIG. 201.

[1001]FIG. 203 shows a nucleotide sequence (SEQ ID NO:482) of a nativesequence PRO1083 cDNA, wherein SEQ ID NO:482 is a clone designatedherein as “UNQ540” and/or “DNA50921-1458”.

[1002]FIG. 204 shows the amino acid sequence (SEQ ID NO:483) derivedfrom the coding sequence of SEQ ID NO:482 shown in FIG. 203.

[1003]FIG. 205 shows an EST nucleotide sequence designated herein asDNA24256 (SEQ ID NO:484).

[1004]FIG. 206 shows a nucleotide sequence (SEQ ID NO:487) of a nativesequence PRO200 cDNA, wherein SEQ ID NO:487 is a clone designated hereinas “UNQ174” and/or “DNA29101-1122”.

[1005]FIG. 207 shows the amino acid sequence (SEQ ID NO:488) derivedfrom the coding sequence of SEQ ID NO:487 shown in FIG. 206.

[1006]FIG. 208 shows a nucleotide sequence (SEQ ID NO:495) of a nativesequence PRO285 cDNA, wherein SEQ ID NO:495 is a clone designated hereinas “DNA40021-1154”.

[1007]FIG. 209 shows the amino acid sequence (SEQ ID NO:496) derivedfrom the coding sequence of SEQ ID NO:495 shown in FIG. 208.

[1008]FIG. 210 shows a nucleotide sequence (SEQ ID NO:497) of a nativesequence PRO286 cDNA, wherein SEQ ID NO:497 is a clone designated hereinas “DNA42663-1154”.

[1009]FIG. 211 shows the amino acid sequence (SEQ ID NO:498) derivedfrom the coding sequence of SEQ ID NO:497 shown in FIG. 210.

[1010]FIG. 212 shows a nucleotide sequence (SEQ ID NO:505) of a nativesequence PRO213-1 cDNA, wherein SEQ ID NO:505 is a clone designatedherein as “DNA30943-1-1163-1”.

[1011]FIG. 213 shows the amino acid sequence (SEQ ID NO:506) derivedfrom the coding sequence of SEQ ID NO:505 shown in FIG. 212.

[1012]FIG. 214 shows a nucleotide sequence (SEQ ID NO:507) of a nativesequence PRO1330 cDNA, wherein SEQ ID NO:507 is a clone designatedherein as “DNA64907-1163-1n.

[1013]FIG. 215 shows the amino acid sequence (SEQ ID NO:508) derivedfrom the coding sequence of SEQ ID NO:507 shown in FIG. 214.

[1014]FIG. 216 shows a nucleotide sequence (SEQ ID NO:509) of a nativesequence PRO1449 cDNA, wherein SEQ ID NO:509 is a clone designatedherein as “DNA64908-1163-1”.

[1015]FIG. 217 shows the amino acid sequence (SEQ ID NO:510) derivedfromthe coding sequence of SEQ ID NO:509 shown in FIG. 216.

[1016]FIG. 218 shows a nucleotide sequence (SEQ ID NO:514) of a nativesequence PRO298 cDNA, wherein SEQ ID NO:514 is a clone designated hereinas “UNQ261” and/or “DNA39975-1210”.

[1017]FIG. 219 shows the amino acid sequence (SEQ ID NO:515) derivedfrom the coding sequence of SEQ ID NO:514 shown in FIG. 218.

[1018]FIG. 220 shows an EST nucleotide sequence designated herein asDNA26832 (SEQ ID NO:516).

[1019]FIG. 221 shows a nucleotide sequence (SEQ ID NO:522) of a nativesequence PRO337 cDNA, wherein SEQ ID NO:522 is a clone designated hereinas “DNA43316-1237”.

[1020]FIG. 222 shows the amino acid sequence (SEQ ID NO:523) derivedfrom the coding sequence of SEQ ID NO:522 shown in FIG. 221.

[1021]FIG. 223 shows an EST nucleotide sequence designated herein asDNA42301 (SEQ ID NO:524).

[1022]FIG. 224 shows a nucleotide sequence (SEQ ID NO:525) of a nativesequence PRO403 cDNA, wherein SEQ ID NO:525 is a clone designated hereinas “DNA55800-1263”.

[1023]FIG. 225 shows the amino acid sequence (SEQ ID NO:526) derivedfrom the coding sequence of SEQ ID NO:525 shown in FIG. 224.

[1024]FIG. 226 shows an EST nucleotide sequence designated herein asDNA34415 (SEQ ID NO:527).

[1025]FIG. 227 shows an EST nucleotide sequence designated herein asDNA49830 (SEQ ID NO:528).

[1026]FIG. 228 shows an EST nucleotide sequence designated herein asDNA49831 (SEQ ID NO:529).

[1027]FIG. 229 shows a nucleotide sequence (SEQ ID NO:611) of a nativesequence PRO4993 cDNA, wherein SEQ ID NO:611 is a clone designatedherein as “DNA94832-2659”.

[1028]FIG. 230 shows the amino acid sequence (SEQ ID NO:612) derivedfrom the coding sequence of SEQ ID NO:611 shown in FIG. 229.

[1029]FIG. 231 shows a nucleotide sequence (SEQ ID NO:613) of a nativesequence PRO1559 cDNA, wherein SEQ ID NO:613 is a clone designatedherein as “DNA68886”.

[1030]FIG. 232 shows the amino acid sequence (SEQ ID NO:614) derivedfrom the coding sequence of SEQ ID NO:613 shown in FIG. 231.

[1031]FIG. 233 shows a nucleotide sequence (SEQ ID NO:615) of a nativesequence PRO725 cDNA, wherein SEQ ID NO:615 is a clone designated hereinas “DNA52758-1399”.

[1032]FIG. 234 shows the amino acid sequence (SEQ ID NO:616) derivedfrom the coding sequence of SEQ ID NO:615 shown in FIG. 233.

[1033]FIG. 235 shows a nucleotide sequence (SEQ ID NO:617) of a nativesequence PRO739 cDNA, wherein SEQ ED NO:617 is a clone designated hereinas “DNA52756”.

[1034]FIG. 236 shows the amino acid sequence (SEQ ID NO:618) derivedfrom the coding sequence of SEQ ID NO:617 shown in FIG. 235.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[1035] I. Definitions

[1036] The terms “PRO polypeptide” and “PRO” as used herein and whenimmediately followed by a numerical designation refer to variouspolypeptides, wherein the complete designation (i.e., PRO/number) refersto specific polypeptide sequences as described herein. The terms“PRO/number polypeptide” and “PRO/number” wherein the term “number” isprovided as an actual numerical designation as used herein encompassnative sequence polypeptides and polypeptide variants (which are furtherdefined herein). The PRO polypeptides described herein may be isolatedfrom a variety of sources, such as from human tissue types or fromanother source, or prepared by recombinant or synthetic methods.

[1037] A “native sequence PRO polypeptide” comprises a polypeptidehaving the same amino acid sequence as the corresponding PRO polypeptidederived from nature. Such native sequence PRO polypeptides can beisolated from nature or can be produced by recombinant or syntheticmeans. The term “native sequence PRO polypeptide” specificallyencompasses naturally-occurring truncated or secreted forms of thespecific PRO polypeptide (e.g., an extracellular domain sequence),naturally-occurring variant forms (e.g., alternatively spliced forms)and naturally-occurring allelic variants of the polypeptide. In variousembodiments of the invention, the native sequence PRO polypeptidesdisclosed herein are mature or full-length native sequence polypeptidescomprising the full-length amino acids sequences shown in theaccompanying figures. Start and stop codons are shown in bold font andunderlined in the figures. However, while the PRO polypeptide disclosedin the accompanying figures are shown to begin with methionine residuesdesignated herein as amino acid position 1 in the figures, it isconceivable and possible that other methionine residues located eitherupstream or downstream from the amino acid position 1 in the figures maybe employed as the starting amino acid residue for the PRO polypeptides.

[1038] The PRO polypeptide “extracellular domain” or “ECD” refers to aform of the PRO polypeptide which is essentially free of thetransmembrane and cytoplasmic domains. Ordinarily, a PRO polypeptide ECDwill have less than 1% of such transmembrane and/or cytoplasmic domainsand preferably, will have less than 0.5% of such domains. It will beunderstood that any transmembrane domains identified for the PROpolypeptides of the present invention are identified pursuant tocriteria routinely employed in the art for identifying that type ofhydrophobic domain. The exact boundaries of a transmembrane domain mayvary but most likely by no more than about 5 amino acids at either endof the domain as initially identified herein. Optionally, therefore, anextracellular domain of a PRO polypeptide may contain from about 5 orfewer amino acids on either side of the transmembranedomain/extracellular domain boundary as identified in the Examples orspecification and such polypeptides, with or without the associatedsignal peptide, and nucleic acid encoding them, are comtemplated by thepresent invention.

[1039] The approximate location of the 'signal peptides” of the variousPRO polypeptides disclosed herein are shown in the present specificationand/or the accompanying figures. It is noted, however, that theC-terminal boundary of a signal peptide may vary, but most likely by nomore than about 5 amino acids on either side of the signal peptideC-terminal boundary as initially identified herein, wherein theC-terminal boundary of the signal peptide may be identified pursuant tocriteria routinely employed in the art for identifying that type ofamino acid sequence element (e.g., Nielsen et al., Prot. Eng. 10:1-6(1997) and von Heinje et al., Nucl. Acids. Res. 14:4683-4690 (1986)).Moreover, it is also recognized that, in some cases, cleavage of asignal sequence from a secreted polypeptide is not entirely uniform,resulting in more than one secreted species. These mature polypeptides,where the signal peptide is cleaved within no more than about 5 aminoacids on either side of the C-terminal boundary of the signal peptide asidentified herein, and the polynucleotides encoding them, arecontemplated by the present invention. “PRO polypeptide variant” meansan active PRO polypeptide as defined above or below having at leastabout 80% amino acid sequence identity with a full-length nativesequence PRO polypeptide sequence as disclosed herein, a PRO polypeptidesequence lacking the signal peptide as disclosed herein, anextracellular domain of a PRO polypeptide, with or without the signalpeptide, as disclosed herein or any other fragment of a full-length PROpolypeptide sequence as disclosed herein. Such PRO polypeptide variantsinclude, for instance, PRO polypeptides wherein one or more amino acidresidues are added, or deleted, at the N- or C-terminus of thefull-length native amino acid sequence. Ordinarily, a PRO polypeptidevariant will have at least about 80% amino acid sequence identity,preferably at least about 81% amino acid sequence identity, morepreferably at least about 82% amino acid sequence identity, morepreferably at least about 83% amino acid sequence identity, morepreferably at least about 84% amino acid sequence identity, morepreferably at least about 85% amino acid sequence identity, morepreferably at least about 86% amino acid sequence identity, morepreferably at least about 87% amino acid sequence identity, morepreferably at least about 88% amino acid sequence identity, morepreferably at least about 89% amino acid sequence identity, morepreferably at least about 90% amino acid sequence identity, morepreferably at least about 91% amino acid sequence identity, morepreferably at least about 92% amino acid sequence identity, morepreferably at least about 93% amino acid sequence identity, morepreferably at least about 94% amino acid sequence identity, morepreferably at least about 95% amino acid sequence identity, morepreferably at least about 96% amino acid sequence identity, morepreferably at least about 97% amino acid sequence identity, morepreferably at least about 98% amino acid sequence identity and mostpreferably at least about 99% amino acid sequence identity with afull-length native sequence PRO polypeptide sequence as disclosedherein, a PRO polypeptide sequence lacking the signal peptide asdisclosed herein, an extracellular domain of a PRO polypeptide, with orwithout the signal peptide, as disclosed herein or any otherspecifically defined fragment of a full-length PRO polypeptide sequenceas disclosed herein. Ordinarily, PRO variant polypeptides are at leastabout l amino acids in length, often at least about 20 amino acids inlength, more often at least about 30 amino acids in length, more oftenat least about 40 amino acids in length, more often at least about 50amino acids in length, more often at least about 60 amino acids inlength, more often at least about 70 amino acids in length, more oftenat least about 80 amino acids in length, more often at least about 90amino acids in length, more often at least about 100 amino acids inlength, more often at least about 150 amino acids in length, more oftenat least about 200 amino acids in length, more often at least about 300amino acids in length, or more. “Percent (%) amino acid sequenceidentity” with respect to the PRO polypeptide sequences identifiedherein is defined as the percentage of amino acid residues in acandidate sequence that are identical with the amino acid residues inthe specific PRO polypeptide sequence, after aligning the sequences andintroducing gaps, if necessary, to achieve the maximum percent sequenceidentity, and not considering any conservative substitutions as part ofthe sequence identity. Alignment for purposes of determining percentamino acid sequence identity can be achieved in various ways that arewithin the skill in the art, for instance, using publicly availablecomputer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR)software. Those skilled in the art can determine appropriate parametersfor measuring alignment, including any algorithms needed to achievemaximal alignment over the full length of the sequences being compared.For purposes herein, however, % amino acid sequence identity values aregenerated using the sequence comparison computer program ALIGN-2,wherein the complete source code for the ALIGN-2 program is provided inTable 1 below. The ALIGN-2 sequence comparison computer program wasauthored by Genentech, Inc. and the source code shown in Table 1 belowhas been filed with user documentation in the U.S. Copyright Office,Washington D.C., 20559, where it is registered under U.S. CopyrightRegistration No. TXU510087. The ALIGN-2 program is publicly availablethrough Genentech, Inc., South San Francisco, Calif. or may be compiledfrom the source code provided in Table 1 below. The ALIGN-2 programshould be compiled for use on a UNIX operating system, preferablydigital UNIX V4.0D. All sequence comparison parameters are set by theALIGN-2 program and do not vary.

[1040] In situations where ALIGN-2 is employed for amino acid sequencecomparisons, the % amino acid sequence identity of a given amino acidsequence A to, with, or against a given amino acid sequence B (which canalternatively be phrased as a given amino acid sequence A that has orcomprises a certain % amino acid sequence identity to, with, or againsta given amino acid sequence B) is calculated as follows:

100 times the fraction X[Y

[1041] where X is the number of amino acid residues scored as identicalmatches by the sequence alignment program ALIGN-2 in that program'salignment of A and B, and where Y is the total number of amino acidresidues in B. It will be appreciated that where the length of aminoacid sequence A is not equal to the length of amino acid sequence B, the% amino acid sequence identity of A to B will not equal the % amino acidsequence identity of B to A. As examples of % amino acid sequenceidentity calculations using this method, Tables 2 and 3 demonstrate howto calculate the % amino acid sequence identity of the amino acidsequence designated “Comparison Protein” to the amino acid sequencedesignated “PRO”, wherein “PRO” represents the amino acid sequence of ahypothetical PRO polypeptide of interest, “Comparison Protein”represents the amino acid sequence of a polypeptide against which the“PRO” polypeptide of interest is being compared, and “X, “Y” and “Z”each represent different hypothetical amino acid residues.

[1042] Unless specifically stated otherwise, all % amino acid sequenceidentity values used herein are obtained as described in the immediatelypreceding paragraph using the ALIGN-2 computer program. However, % aminoacid sequence identity values may also be obtained as described below byusing the WU-BLAST-2 computer program (Altschul et al., Methods inEnzymology 266:460-480 (1996)). Most of the WU-BLAST-2 search parametersare set to the default values. Those not set to default values, i.e.,the adjustable parameters, are set with the following values: overlapspan=1, overlap fraction=0.125, word threshold (T)=11, and scoringmatrix=BLOSUM62. When WU-BLAST-2 is employed, a % amino acid sequenceidentity value is determined by dividing (a) the number of matchingidentical amino acid residues between the amino acid sequence of the PROpolypeptide of interest having a sequence derived from the native PROpolypeptide and the comparison amino acid sequence of interest (i.e.,the sequence against which the PRO polypeptide of interest is beingcompared which may be a PRO variant polypeptide) as determined byWU-BLAST-2 by (b) the total number of amino acid residues of the PROpolypeptide of interest. For example, in the statement “a polypeptidecomprising an the amino acid sequence A which has or having at least 80%amino acid sequence identity to the amino acid sequence B”, the aminoacid sequence A is the comparison amino acid sequence of interest andthe amino acid sequence B is the amino acid sequence of the PROpolypeptide of interest.

[1043] Percent amino acid sequence identity may also be determined usingthe sequence comparison program NCBI-BLAST2 (Altschul et al., NucleicAcids Res. 25:3389-3402 (1997)). The NCBI-BLAST2 sequence comparisonprogram may be downloaded from http://www.ncbi.nhn.nih.gov. NCBI-BLAST2uses several search parameters, wherein all of those search parametersare set to default values including, for example, unmask=yes,strand=all, expected occurrences=10, minimum low complexity length=15/5,multi-pass e-value=0.01, constant for multi-pass=25, dropoff for finalgapped alignment=25 and scoring matrix=BLOSUM62.

[1044] In situations where NCBI-BLAST2 is employed for amino acidsequence comparisons, the % amino acid sequence identity of a givenamino acid sequence A to, with, or against a given amino acid sequence B(which can alternatively be phrased as a given amino acid sequence Athat has or comprises a certain % amino acid sequence identity to, with,or against a given amino acid sequence B) is calculated as follows:

100 times the fraction X[Y

[1045] where X is the number of amino acid residues scored as identicalmatches by the sequence alignment program NCBI-BLAST2 in that program'salignment of A and B, and where Y is the total number of amino acidresidues in B. It will be appreciated that where the length of aminoacid sequence A is not equal to the length of amino acid sequence B, the% amino acid sequence identity of A to B will not equal the % amino acidsequence identity of B to A. “PRO variant polynucleotide” or “PROvariant nucleic acid sequence” means a nucleic acid molecule whichencodes an active PRO polypeptide as defined below and which has atleast about 80% nucleic acid sequence identity with a nucleotide acidsequence encoding a full-length native sequence PRO polypeptide sequenceas disclosed herein, a full-length native sequence PRO polypeptidesequence lacking the signal peptide as disclosed herein, anextracellular domain of a PRO polypeptide, with or without the signalpeptide, as disclosed herein or any other fragment of a full-length PROpolypeptide sequence as disclosed herein. Ordinarily, a PRO variantpolynucleotide will have at least about 80% nucleic acid sequenceidentity, more preferably at least about 81% nucleic acid sequenceidentity, more preferably at least about 82% nucleic acid sequenceidentity, more preferably at least about 83% nucleic acid sequenceidentity, more preferably at least about 84% nucleic acid sequenceidentity, more preferably at least about 85% nucleic acid sequenceidentity, more preferably at least about 86% nucleic acid sequenceidentity, more preferably at least about 87% nucleic acid sequenceidentity, more preferably at least about 88% nucleic acid sequenceidentity, more preferably at least about 89% nucleic acid sequenceidentity, more preferably at least about 90% nucleic acid sequenceidentity, more preferably at least about 91% nucleic acid sequenceidentity, more preferably at least about 92% nucleic acid sequenceidentity, more preferably at least about 93% nucleic acid sequenceidentity, more preferably at least about 94% nucleic acid sequenceidentity, more preferably at least about 95% nucleic acid sequenceidentity, more preferably at least about 96% nucleic acid sequenceidentity, more preferably at least about 97% nucleic acid sequenceidentity, more preferably at least about 98% nucleic acid sequenceidentity and yet more preferably at least about 99% nucleic acidsequence identity with a nucleic acid sequence encoding a full-lengthnative sequence PRO polypeptide sequence as disclosed herein, afull-length native sequence PRO polypeptide sequence lacking the signalpeptide as disclosed herein, an extracellular domain of a PROpolypeptide, with or without the signal sequence, as disclosed herein orany other fragment of a full-length PRO polypeptide sequence asdisclosed herein. Variants do not encompass the native nucleotidesequence.

[1046] Ordinarily, PRO variant polynucleotides are at least about 30nucleotides in length, often at least about 60 nucleotides in length,more often at least about 90 nucleotides in length, more often at leastabout 120 nucleotides in length, more often at least about 150nucleotides in length, more often at least about 180 nucleotides inlength, more often at least about 210 nucleotides in length, more oftenat least about 240 nucleotides in length, more often at least about 270nucleotides in length, more often at least about 300 nucleotides inlength, more often at least about 450 nucleotides in length, more oftenat least about 600 nucleotides in length, more often at least about 900nucleotides in length, or more.

[1047] “Percent (%) nucleic acid sequence identity” with respect toPRO-encoding nucleic acid sequences identified herein is defined as thepercentage of nucleotides in a candidate sequence that are identicalwith the nucleotides in the PRO nucleic acid sequence of interest, afteraligning the sequences and introducing gaps, if necessary, to achievethe maximum percent sequence identity. Alignment for purposes ofdetermining percent nucleic acid sequence identity can be achieved invarious ways that are within the skill in the art, for instance, usingpublicly available computer software such as BLAST, BLAST-2, ALIGN orMegalign (DNASTAR) software. For purposes herein, however, % nucleicacid sequence identity values are generated using the sequencecomparison computer program ALIGN-2, wherein the complete source codefor the ALIGN-2 program is provided in Table 1 below. The ALIGN-2sequence comparison computer program was authored by Genentech, Inc. andthe source code shown in Table 1 below has been filed with userdocumentation in the U.S. Copyright Office, Washington D.C., 20559,where it is registered under U.S. Copyright Registration No. TXU510087.The ALIGN-2 program is publicly available through Genentech, Inc., SouthSan Francisco, Calif. or may be compiled from the source code providedin Table 1 below. The ALIGN-2 program should be compiled for use on aUNIX operating system, preferably digital UNIX V4.0D. All sequencecomparison parameters are set by the ALIGN-2 program and do not vary.

[1048] In situations where ALIGN-2 is employed for nucleic acid sequencecomparisons, the % nucleic acid sequence identity of a given nucleicacid sequence C to, with, or against a given nucleic acid sequence D(which can alternatively be phrased as a given nucleic acid sequence Cthat has or comprises a certain % nucleic acid sequence identity to,with, or against a given nucleic acid sequence D) is calculated asfollows:

100 times the fraction W/Z

[1049] where W is the number of nucleotides scored as identical matchesby the sequence alignment program ALIGN-2 in that program's alignment ofC and D, and where Z is the total number of nucleotides in D. It will beappreciated that where the length of nucleic acid sequence C is notequal to the length of nucleic acid sequence D, the % nucleic acidsequence identity of C to D will not equal the % nucleic acid sequenceidentity of D to C. As examples of % nucleic acid sequence identitycalculations, Tables 4 and 5, demonstrate how to calculate the % nucleicacid sequence identity of the nucleic acid sequence designated“Comparison DNA” to the nucleic acid sequence designated “PRO-DNA”,wherein “PRO-DNA” represents a hypothetical PRO-encoding nucleic acidsequence of interest, “Comparison DNA” represents the nucleotidesequence of a nucleic acid molecule against which the “PRO-DNA” nucleicacid molecule of interest is being compared, and “N”, “L” and “V” eachrepresent different hypothetical nucleotides.

[1050] Unless specifically stated otherwise, all % nucleic acid sequenceidentity values used herein are obtained as described in the immediatelypreceding paragraph using the ALIGN-2 computer program. However, %nucleic acid sequence identity values may also be obtained as describedbelow by using the WU-BLAST-2 computer program (Altschul et al., Methodsin Enzymology 266:460-480 (1996)). Most of the WU-BLAST-2 searchparameters are set to the default values. Those not set to defaultvalues, i.e., the adjustable parameters, are set with the followingvalues: overlap span=1, overlap fraction=0.125, word threshold (T)=11,and scoring matrix=BLOSUM62. When WU-BLAST-2 is employed, a % nucleicacid sequence identity value is determined by dividing (a) the number ofmatching identical nucleotides between the nucleic acid sequence of thePRO polypeptide-encoding nucleic acid molecule of interest having asequence derived from the native sequence PRO polypeptide-encodingnucleic acid and the comparison nucleic acid molecule of interest (i.e.,the sequence against which the PRO polypeptide-encoding nucleic acidmolecule of interest is being compared which may be a variant PROpolynucleotide) as determined by WU-BLAST-2 by (b) the total number ofnucleotides of the PRO polypeptide-encoding nucleic acid molecule ofinterest. For example, in the statement “an isolated nucleic acidmolecule comprising a nucleic acid sequence A which has or having atleast 80% nucleic acid sequence identity to the nucleic acid sequenceB”, the nucleic acid sequence A is the comparison nucleic acid moleculeof interest and the nucleic acid sequence B is the nucleic acid sequenceof the PRO polypeptide-encoding nucleic acid molecule of interest.

[1051] Percent nucleic acid sequence identity may also be determinedusing the sequence comparison program NCBI-BLAST2 (Altschul et al.,Nucleic Acids Res. 25:3389-3402 (1997)). The NCBI-BLAST2 sequencecomparison program may be downloaded from http://www.ncbi.nlm.nih.gov.NCBI-BLAST2 uses several search parameters, wherein all of those searchparameters are set to default values including, for example, unmask=yes,strand=all, expected occurrences=10, minimum low complexity length=15/5,multi-pass e-value=0.01, constant for multi-pass=25, dropoff for finalgapped alignment=25 and scoring matrix=BLOSUM62.

[1052] In situations where NCBI-BLAST2 is employed for sequencecomparisons, the % nucleic acid sequence identity of a given nucleicacid sequence C to, with, or against a given nucleic acid sequence D(which can alternatively be phrased as a given nucleic acid sequence Cthat has or comprises a certain % nucleic acid sequence identity to,with, or against a given nucleic acid sequence D) is calculated asfollows:

100 times the fraction W/Z

[1053] where W is the number of nucleotides scored as identical matchesby the sequence alignment program NCBI-BLAST2 in that program'salignment of C and D, and where Z is the total number of nucleotides inD. It will be appreciated that where the length of nucleic acid sequenceC is not equal to the length of nucleic acid sequence D, the % nucleicacid sequence identity of C to D will not equal the % nucleic acidsequence identity of D to C.

[1054] In other embodiments, PRO variant polynucleotides are nucleicacid molecules that encode an active PRO polypeptide and which arecapable of hybridizing, preferably under stringent hybridization andwash conditions, to nucleotide sequences encoding a full-length PROpolypeptide as disclosed herein. PRO variant polypeptides may be thosethat are encoded by a PRO variant polynucleotide.

[1055] The term “positives”, in the context of sequence comparisonperformed as described above, includes residues in the sequencescompared that are not identical but have similar properties (e.g. as aresult of conservative substitutions, see Table 6 below). For purposesherein, the % value of positives is determined by dividing (a) thenumber of amino acid residues scoring a positive value between the PROpolypeptide amino acid sequence of interest having a sequence derivedfrom the native PRO polypeptide sequence and the comparison amino acidsequence of interest (i.e., the amino acid sequence against which thePRO polypeptide sequence is being compared) as determined in theBLOSUM62 matrix of WU-BLAST-2 by (b) the total number of amino acidresidues of the PRO polypeptide of interest.

[1056] Unless specifically stated otherwise, the % value of positives iscalculated as described in the immediately preceding paragraph. However,in the context of the amino acid sequence identity comparisons performedas described for ALIGN-2 and NCBI-BLAST-2 above, includes amino acidresidues in the sequences compared that are not only identical, but alsothose that have similar properties. Amino acid residues that score apositive value to an amino acid residue of interest are those that areeither identical to the amino acid residue of interest or are apreferred substitution (as defined in Table 6 below) of the amino acidresidue of interest.

[1057] For amino acid sequence comparisons using ALIGN-2 or NCBI-BLAST2,the % value of positives of a given amino acid sequence A to, with, oragainst a given amino acid sequence B (which can alternatively bephrased as a given amino acid sequence A that has or comprises a certain% positives to, with, or against a given amino acid sequence B) iscalculated as follows:

100 times the fraction X/Y

[1058] where X is the number of amino acid residues scoring a positivevalue as defined above by the sequence alignment program ALIGN-2 orNCBI-BLAST2 in that program's alignment of A and B, and where Y is thetotal number of amino acid residues in B. It will be appreciated thatwhere the length of amino acid sequence A is not equal to the length ofamino acid sequence B, the % positives of A to B will not equal the %positives of B to A.

[1059] “Isolated,” when used to describe the various polypeptidesdisclosed herein, means polypeptide that has been identified andseparated and/or recovered from a component of its natural environment.Contaminant components of its natural environment are materials thatwould typically interfere with diagnostic or therapeutic uses for thepolypeptide, and may include enzymes, hormones, and other proteinaceousor non-proteinaceous solutes. In preferred embodiments, the polypeptidewill be purified (1) to a degree sufficient to obtain at least 15residues of N-terminal or internal amino acid sequence by use of aspinning cup sequenator, or (2) to homogeneity by SDS-PAGE undernon-reducing or reducing conditions using Coomassie blue or, preferably,silver stain. Isolated polypeptide includes polypeptide in situ withinrecombinant cells, since at least one component of the PRO polypeptidenatural environment will not be present. Ordinarily, however, isolatedpolypeptide will be prepared by at least one purification step.

[1060] An “isolated” PRO polypeptide-encoding nucleic acid or otherpolypeptide-encoding nucleic acid is a nucleic acid molecule that isidentified and separated from at least one contaminant nucleic acidmolecule with which it is ordinarily associated in the natural source ofthe polypeptide-encoding nucleic acid. An isolated polypeptide-encodingnucleic acid molecule is other than in the form or setting in which itis found in nature. Isolated polypeptide-encoding nucleic acid moleculestherefore are distinguished from the specific polypeptide-encodingnucleic acid molecule as it exists in natural cells. However, anisolated polypeptide-encoding nucleic acid molecule includespolypeptide-encoding nucleic acid molecules contained in cells thatordinarily express the polypeptide where, for example, the nucleic acidmolecule is in a chromosomal location different from that of naturalcells.

[1061] The term “control sequences” refers to DNA sequences necessaryfor the expression of an operably linked coding sequence in a particularhost organism. The control sequences that are suitable for prokaryotes,for example, include a promoter, optionally an operator sequence, and aribosome binding site. Eukaryotic cells are known to utilize promoters,polyadenylation signals, and enhancers.

[1062] Nucleic acid is “operably linked” when it is placed into afunctional relationship with another nucleic acid sequence. For example,DNA for a presequence or secretory leader is operably linked to DNA fora polypeptide if it is expressed as a preprotein that participates inthe secretion of the polypeptide; a promoter or enhancer is operablylinked to a coding sequence if it affects the transcription of thesequence; or a ribosome binding site is operably linked to a codingsequence if it is positioned so as to facilitate translation. Generally,“operably linked” means that the DNA sequences being linked arecontiguous, and, in the case of a secretory leader, contiguous and inreading phase. However, enhancers do not have to be contiguous. Linkingis accomplished by ligation at convenient restriction sites. If suchsites do not exist, the synthetic oligonucleotide adaptors or linkersare used in accordance with conventional practice.

[1063] The term “antibody” is used inthe broadest sense and specificallycovers, for example, single anti-PRO monoclonal antibodies (includingagonist, antagonist, and neutralizing antibodies), anti-PRO antibodycompositions with polyepitopic specificity, single chain anti-PROantibodies, and fragments of anti-PRO antibodies (see below). The term“monoclonal antibody” as used herein refers to an antibody obtained froma population of substantially homogeneous antibodies, i.e., theindividual antibodies comprising the population are identical except forpossible naturally-occurring mutations that may be present in minoramounts.

[1064] “Stringency” of hybridization reactions is readily determinableby one of ordinary skill in the art, and generally is an empiricalcalculation dependent upon probe length, washing temperature, and saltconcentration. In general, longer probes require higher temperatures forproper annealing, while shorter probes need lower temperatures.Hybridization generally depends on the ability of denatured DNA toreanneal when complementary strands are present in an environment belowtheir melting temperature. The higher the degree of desired homologybetween the probe and hybridizable sequence, the higher the relativetemperature which can be used. As a result, it follows that higherrelative temperatures would tend to make the reaction conditions morestringent, while lower temperatures less so. For additional details andexplanation of stringency of hybridization reactions, see Ausubel etal., Current Protocols in Molecular Biology, Wiley IntersciencePublishers, (1995). “Stringent conditions” or “high stringencyconditions”, as defined herein, may be identified by those that: (1)employ low ionic strength and high temperature for washing, for example0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecylsulfate at 50° C.; (2) employ during hybridization a denaturing agent,such as formamide, for example, 50% (v/v) formamide with 0.1% bovineserum albumin/0.1% Ficoll/0.1% polyvinylpyrrolidone/50 mM sodiumphosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodiumcitrate at 42° C.; or (3) employ 50% formamide, 5× SSC (0.75 M NaCl,0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodiumpyrophosphate, 5× Denhardt's solution, sonicated salmon sperm DNA (50μg/ml), 0.1% SDS, and 10% dextran sulfate at 42° C., with washes at 42°C. in 0.2× SSC (sodium chloride/sodium citrate) and 50% formamide at 55°C., followed by a high-stringency wash consisting of 0.1× SSC containingEDTA at 55° C. “Moderately stringent conditions” may be identified asdescribed by Sambrook et al., Molecular Cloning: A Laboratory Manual,New York: Cold Spring Harbor Press, 1989, and include the use of washingsolution and hybridization conditions (e.g., temperature, ionic strengthand %SDS) less stringent that those described above. An example ofmoderately stringent conditions is overnight incubation at 37° C. in asolution comprising: 20% formamide, 5× SSC (150 mM NaCl, 15 mM trisodiumcitrate), 50 mM sodium phosphate (pH 7.6), 5× Denhardt's solution, 10%dextran sulfate, and 20 mg/ml denatured sheared salmon sperm DNA,followed by washing the filters in 1× SSC at about 37-50° C. The skilledartisan will recognize how to adjust the temperature, ionic strength,etc. as necessary to accommodate factors such as probe length and thelike.

[1065] The term “epitope tagged” when used herein refers to a chimericpolypeptide comprising a PRO polypeptide fused to a “tag polypeptide”.The tag polypeptide has enough residues to provide an epitope againstwhich an antibody can be made, yet is short enough such that it does notinterfere with activity of the polypeptide to which it is fused. The tagpolypeptide preferably also is fairly unique so that the antibody doesnot substantially cross-react with other epitopes. Suitable tagpolypeptides generally have at least six amino acid residues and usuallybetween about 8 and 50 amino acid residues (preferably, between about 10and 20 amino acid residues).

[1066] As used herein, the term “immunoadhesin” designates antibody-likemolecules which combine the binding specificity of a heterologousprotein (an “adhesin”) with the effector functions of immunoglobulinconstant domains. Structurally, the immunoadhesins comprise a fusion ofan amino acid sequence with the desired binding specificity which isother than the antigen recognition and binding site of an antibody(i.e., is “heterologous”), and an immunoglobulin constant domainsequence. The adhesin part of an immunoadhesin * molecule typically is acontiguous amino acid sequence comprising at least the binding site of areceptor or a ligand. The immunoglobulin constant domain sequence in theimmunoadhesin may be obtained from any immunoglobulin, such as IgG-1,IgG-2, IgG-3, or IgG4 subtypes, IgA (including IgA-1 and IgA-2), IgE,IgD or IgM.

[1067] “Active” or “activity” for the purposes herein refers to form(s)of a PRO polypeptide which retain a biological and/or an immunologicalactivity of native or naturally-occurring PRO, wherein “biological”activity refers to a biological function (either inhibitory orstimulatory) caused by a native or naturally-occurring PRO other thanthe ability to induce the production of an antibody against an antigenicepitope possessed by a native or naturally-occurring PRO and an“immunological” activity refers to the ability to induce the productionof an antibody against an antigenic epitope possessed by a native ornaturally-occurring PRO.

[1068] The term “antagonist” is used in the broadest sense, and includesany molecule that partially or fully blocks, inhibits, or neutralizes abiological activity of a native PRO polypeptide disclosed herein. In asimilar manner, the term “agonist” is used in the broadest sense andincludes any molecule that mimics a biological activity of a native PROpolypeptide disclosed herein. Suitable agonist or antagonist moleculesspecifically include agonist or antagonist antibodies or antibodyfragments, fragments or amino acid sequence variants of native PROpolypeptides, peptides, antisense oligonucleotides, small organicmolecules, etc. Methods for identifying agonists or antagonists of a PROpolypeptide may comprise contacting a PRO polypeptide with a candidateagonist or antagonist molecule and measuring a detectable change in oneor more biological activities normally associated with the PROpolypeptide.

[1069] “Treatment” refers to both therapeutic treatment and prophylacticor preventative measures, wherein the object is to prevent or slow down(lessen) the targeted pathologic condition or disorder. Those in need oftreatment include those already with the disorder as well as those proneto have the disorder or those in whom the disorder is to be prevented.

[1070] “Chronic” administration refers to administration of the agent(s)in a continuous mode as opposed to an acute mode, so as to maintain theinitial therapeutic effect (activity) for an extended period of time.“Intermittent” administration is treatment that is not consecutivelydone without interruption, but rather is cyclic in nature.

[1071] “Mammal” for purposes of treatment refers to any animalclassified as a mammal, including humans, domestic and farm animals, andzoo, sports, or pet animals, such as dogs, cats, cattle, horses, sheep,pigs, goats, rabbits, etc. Preferably, the mammal is human.

[1072] Administration “in combination with” one or more furthertherapeutic agents includes simultaneous (concurrent) and consecutiveadministration in any order.

[1073] “Carriers” as used herein include pharmaceutically acceptablecarriers, excipients, or stabilizers which are nontoxic to the cell ormammal being exposed thereto at the dosages and concentrations employed.Often the physiologically acceptable carrier is an aqueous pH bufferedsolution. Examples of physiologically acceptable carriers includebuffers such as phosphate, citrate, and other organic acids;antioxidants including ascorbic acid; low molecular weight (less thanabout 10 residues) polypeptide; proteins, such as serum albumin,gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides, andother carbohydrates including glucose, mamnose, or dextrins; chelatingagents such as EDTA; sugar alcohols such as mannitol or sorbitol;salt-forming counterions such as sodium; and/or nonionic surfactantssuch as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[1074] “Antibody fragments” comprise a portion of an intact antibody,preferably the antigen binding or variable region of the intactantibody. Examples of antibody fragments include Fab, Fab′, F(ab′)₂, andFv fragments; diabodies; linear antibodies (Zapata et al., Protein Eng.8(10):1057-1062 [1995]); single-chain antibody molecules; andmultispecific antibodies formed from antibody fragments.

[1075] Papain digestion of antibodies produces two identicalantigen-binding fragments, called “Fab” fragments, each with a singleantigen-binding site, and a residual “Fc” fragment, a designationreflecting the ability to crystallize readily. Pepsin treatment yieldsan F(ab′)₂ fragment that has two antigen-combining sites and is stillcapable of cross-linking antigen.

[1076] “Fv” is the minimun antibody fragment which contains a completeantigen-recognition and -binding site. This region consists of a dimerof one heavy- and one light-chain variable domain in tight, non-covalentassociation. It is in this configuration that the three CDRs of eachvariable domain interact to define an antigen-binding site on thesurface of the V_(H)-V_(L) dimer. Collectively, the six CDRs conferantigen-binding specificity to the antibody. However, even a singlevariable domain (or half of an Fv comprising only three CDRs specificfor an antigen) has the ability to recognize and bind antigen, althoughat a lower affinity than the entire binding site.

[1077] The Fab fragment also contains the constant domain of the lightchain and the first constant domain (CH1) of the heavy chain. Fabfragments differ from Fab′ fragments by the addition of a few residuesat the carboxy terminus of the heavy chain CH1 domain including one ormore cysteines from the antibody hinge region. Fab′-SH is thedesignation herein for Fab′ in which the cysteine residue(s) of theconstant domains bear a free thiol group. F(ab′)₂ antibody fragmentsoriginally were produced as pairs of Fab′ fragments which have hingecysteines between them. Other chemical couplings of antibody fragmentsare also known.

[1078] The “light chains” of antibodies (immunoglobulins) from anyvertebrate species can be assigned to one of two clearly distinct types,called kappa and lambda, based on the amino acid sequences of theirconstant domains.

[1079] Depending on the amino acid sequence of the constant domain oftheir heavy chains, immunoglobulins can be assigned to differentclasses. There are five major classes of immunoglobulins: IgA, IgD, IgE,IgG, and IgM, and several of these may be further divided intosubclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2.

[1080] “Single-chain Fv” or “sFv” antibody fragments comprise the VH andVL domains of antibody, wherein these domains are present in a singlepolypeptide chain. Preferably, the Fv polypeptide further comprises apolypeptide linker between the V_(H) and V_(L) domains which enables thesFv to form the desired structure for antigen binding. For a review ofsFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol.113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315(1994).

[1081] The term “diabodies” refers to small antibody fragments with twoantigen-binding sites, which fragments comprise a heavy-chain variabledomain (V_(H)) connected to a light-chain variable domain (V_(L)) in thesame polypeptide chain (V_(H)-V_(L)). By using a linker that is tooshort to allow pairing between the two domains on the same chain, thedomains are forced to pair with the complementary domains of anotherchain and create two antigen-binding sites. Diabodies are described morefully in, for example, EP 404,097; WO 93/11161; and Hollinger et al.,Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993).

[1082] An “isolated” antibody is one which has been identified andseparated and/or recovered from a component of its natural environment.Contaminant components of its natural environment are materials whichwould interfere with diagnostic or therapeutic uses for the antibody,and may include enzymes, hormones, and -other proteinaceous ornonproteinaceous solutes. In preferred embodiments, the antibody will bepurified (1) to greater than 95% by weight of antibody as determined bythe Lowry method, and most preferably more than 99% by weight, (2) to adegree sufficient to obtain at least 15 residues of N-terminal orinternal amino acid sequence by use of a spinning cup sequenator, or (3)to homogeneity by SDS-PAGE under reducing or nonreducing conditionsusing Coomassie blue or, preferably, silver stain. Isolated antibodyincludes the antibody in situ within recombinant cells since at leastone component of the antibody's natural environment will not be present.Ordinarily, however, isolated antibody will be prepared by at least onepurification step.

[1083] The word “label” when used herein refers to a detectable compoundor composition which is conjugated directly or indirectly to theantibody so as to generate a “labeled” antibody. The label may bedetectable by itself (e.g. radioisotope labels or fluorescent labels)or, in the case of an enzymatic label, may catalyze chemical alterationof a substrate compound or composition which is detectable.

[1084] By “solid phase” is meant a non-aqueous matrix to which theantibody of the present invention can adhere. Examples of solid phasesencompassed herein include those formed partially or entirely of glass(e.g., controlled pore glass), polysaccharides (e.g., agarose),polyacrylamides, polystyrene, polyvinyl alcohol and silicones. Incertain embodiments, depending on the context, the solid phase cancomprise the well of an assay plate; in others it is a purificationcolumn (e.g., an affinity chromatography column). This term alsoincludes a discontinuous solid phase of discrete particles, such asthose described in U.S. Pat. No. 4,275,149.

[1085] A “liposome” is a small vesicle composed of various types oflipids, phospholipids and/or surfactant which is useful for delivery ofa drug (such as a PRO polypeptide or antibody thereto) to a mammal. Thecomponents of the liposome are commonly arranged in a bilayer formation,similar to the lipid arrangement of biological membranes.

[1086] A 'small molecule” is defined herein to have a molecular weightbelow about 500 Daltons.

[1087] As used herein, “vascular endothelial cell growth factor-E,” or“VEGF-E,” refers to a mammalian growth factor as described herein,including the human amino acid sequence of FIG. 207, a sequence whichhas homology to VEGF and bone morphogenetic protein 1 and which includescomplete conservation of all VEGF cysteine residues, which have beenshown to be required for biological activity of VEGF. VEGF-E expressionincludes expression in human fetal bone, thymus, and thegastrointestinal tract. The biological activity of native VEGF-E isshared by any analogue or variant thereof that is capable of promotingselective growth and/or survival of umbilical vein endothelial cells,induces proliferation of pluripotent fibroblast cells, induces immediateearly gene c-fos in human endothelial cell lines and causes myocytehypertrophy in cardiac cells, or which possesses an immune epitope thatis immunologically cross-reactive with an antibody raised against atleast one epitope of the corresponding native VEGF-E. The human VEGF-Eherein is active on rat and mouse cells indicating conservation acrossspecies. Moreover, the VEGF-E herein is expressed at the growth plateregion and has been shown to embrace fetal myocytes.

[1088] As used herein, “vascular endothelial cell growth factor,” or“VEGF,” refers to a mammalian growth factor as defined in U.S. Pat. No.5,332,671. The biological activity of native VEGF is shared by anyanalogue or variant thereof that is capable of promoting selectivegrowth of vascular endothelial cells but not of bovine cornealendothelial cells, lens epithelial cells, adrenal cortex cells, BHK-21fibroblasts, or keratinocytes, or that possesses an immune epitope thatis immunologically cross-reactive with an antibody raised against atleast one epitope of the corresponding native VEGF.

[1089] The terms “VEGF-E polypeptide” and “VEGF-E” when used hereinencompass native sequence VEGF-E polypeptide and VEGF-E polypeptidevariants (which are further defined herein). The VEGF-E polypeptides maybe isolated from a variety of sources, such as from human tissue typesor from another source, or prepared by recombinant or synthetic methods.

[1090] Inhibitors of VEGF-E include those which reduce or inhibit theactivity or expression of VEGF-E and includes antisense molecules.

[1091] The abbreviation “KDR” refers to the kinase domain region of theVEGF molecule. VEGF-E has no homology with VEGF in this domain.

[1092] The abbreviation “FLT-1” refers to the FMS-like tyrosine kinasebinding domain which is known to bind to the corresponding FLT-1receptor. VEGF-E has no homology with VEGF in this domain.

[1093] “Toll receptor2”, “TLR2” and “huTLR2” are used interchangeably,and refer to a human ToU receptor designated as “HuTLR2” by Rock et al.,Proc. Natl. Acad. Sci. USA 95, 588-593 (1998).

[1094] The term “lipopolysaccharide” or “LPS” is used herein as asynonym of “endotoxin.” Lipopolysaccharides (LPS) are characteristiccomponents of the outer membrane of Gram-negative bacteria, e.g.,Escherichia coli. They consist of a polysaccharide part and a fat calledlipid A. The polysaccharide, which varies from one bacterial species toanother, is made up of the O-specific chain (built from repeating unitsof three to eight sugars) and the two-part core. Lipid A virtuallyalways includes two glucosamine sugars modified by phosphate and avariable number of fatty acids. For further information see, forexample, Rietschel and Brade, Scientific American August 1992, 54-61.

[1095] The term 'septic shock” is used herein in the broadest sense,including all definitions disclosed in Bone, Ann. Intern Med. 114,332-333 (1991). Specifically, septic shock starts with a systemicresponse to infection, a syndrome called sepsis. When this syndromeresults in hypotension and organ dysfunction, it is called septic shock.Septic shock may be initiated by gram-positive organisms and fungi, aswell as endotoxin-containing Gram-negative organisms. Accordingly, thepresent definition is not limited to “endotoxin shock.”

[1096] The phrases “gene amplification” and “gene duplication” are usedinterchangeably and refer to a process by which multiple copies of agene or gene fragment are formed in a particular cell or cell line. Theduplicated region (a stretch of amplified DNA) is often referred to as“amplicon”. Usually, the amount of the messenger RNA (mRNA) produced,i.e., the level of gene expression, also increases in the proportion ofthe number of copies made of the particular gene expressed.

[1097] “Tumor”, as used herein, refers to all neoplastic cell growth andproliferation, whether malignant or benign, and all pre-cancerous andcancerous cells and tissues. The terms “cancer” and “cancerous” refer toor describe the physiological condition in mammals that is typicallycharacterized by unregulated cell growth. Examples of cancer include butare not limited to, carcinoma, lymphoma, blastoma, sarcoma, andleukemia. More particular examples of such cancers include breastcancer, prostate cancer, colon cancer, squamous cell cancer, small-celllung cancer, non-small cell lung cancer, gastrointestinal cancer,pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, livercancer, bladder cancer, hepatoma, colorectal cancer, endometrialcarcinoma, salivary gland carcinoma, kidney cancer, vulval cancer,thyroid cancer, hepatic carcinoma and various types of head and neckcancer.

[1098] The term “cytotoxic agent” as used herein refers to a substancethat inhibits or prevents the function of cells and/or causesdestruction of cells. The term is intended to include radioactiveisotopes (e.g. I131, I_(125,) Y90 and Re186), chemotherapeutic agents,and toxins such as enzymatically active toxins of bacterial, fungal,plant or animal origin, or fragments thereof.

[1099] A “chemotherapeutic agent” is a chemical compound useful in thetreatment of cancer. Examples of chemotherapeutic agents includeadriamycin, doxorubicin, epirubicin, 5-fluorouracil, cytosinearabinoside (“Ara-C”), cyclophosphamide, thiotepa, busulfan, cytoxin,taxoids, e.g. paclitaxel Craxol, Bristol-Myers Squibb Oncology,Princeton, N.J.), and doxetaxel (Taxotere<<, Rhone-Poulenc Rorer,Antony, France), toxotere, methotrexate, cisplatin, melphalan,vinblastine, bleomycin, etoposide, ifosfamide, mitomycin C,mitoxantrone, vincristine, vinorelbine, carboplatin, teniposide,daunomycin, carminomycin, aminopterin, dactinomycin, mitomycins,esperamicins (see U.S. Pat. No. 4,675,187), melphalan and other relatednitrogen mustards. Also included in this definition are hormonal agentsthat act to regulate or inhibit hormone action on tumors such astamoxifen and onapristone.

[1100] A “growth inhibitory agent” when used herein refers to a compoundor composition which inhibits growth of a cell, especially cancer celloverexpressing any of the genes identified herein, either in vitro or invivo. Thus, the growth inhibitory agent is one which significantlyreduces the percentage of cells overexpressing such genes in S phase.Examples of growth inhibitory agents include agents that block cellcycle progression (at a place other than S phase), such as agents thatinduce GI arrest and M-phase arrest. Classical M-phase blockers includethe vincas (vincristine and vinblastine), taxol, and topo II inhibitorssuch as doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin.Those agents that arrest G1 also spill over into S-phase arrest, forexample, DNA alkylating agents such as tamoxifen, prednisone,dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil,and ara-C. Further information can be found in The Molecular Basis ofCancer, Mendelsohn and Israel, eds., Chapter 1, entitled “Cell cycleregulation, oncogens, and antineoplastic drugs” by Murakai et al. (W BSaunders: Philadelphia, 1995), especially p.13.

[1101] “Doxorubicin” is an athracycline antibiotic.

[1102] The term “cytokine” is a generic term for proteins released byone cell population which act on another cell as intercellularmediators. Examples of such cytokines are lymphokines, monokines, andtraditional polypeptide hormones. Included among the cytokines aregrowth hormone such as human growth hormone, N-methionyl human growthhormone, and bovine growth hormone; parathyroid hormone; thyroxine;insulin; proinsulin; relaxin; prorelaxin; and the like. As used herein,the term cytokine includes proteins from natural sources or fromrecombinant cell culture and biologically active equivalents of thenative sequence cytokines. TABLE 1 /*  *  * C-C increased from 12 to 15 * Z is average of EQ  * B is average of ND  * match with stop is _M;stop-stop = 0; J (joker) match = 0  */ #define _M −8 /* value of a matchwith a stop */ int _day[26][26] = { /*  A B C D E F G H I J K L M N O PQ R S T U V W X Y Z */ /* A */ {2, 0, −2, 0, 0, −4, 1, −1, −1, 0, −1,−2, −1, 0, _M, 1, 0, −2, 1, 1, 0, 0, −6, 0, −3, 0}, /* B */ {0, 3, −4,3, 2, −5, 0, 1, −2, 0, 0, −3, −2, 2, _M, −1, 1, 0, 0, 0, 0, −2, −5, 0,−3, 1}, /* C */ {−2, −4, 15, −5, −5, −4, −3, −3, −2, 0, −5, −6, −5, −4,_M, −3, −5, −4, 0, −2, 0, −2, −8, 0, 0, −5}, /* D */ {0, 3, −5, 4, 3,−6, 1, 1, −2, 0, 0, −4, −3, 2, _M, −1, 2, −1, 0, 0, 0, −2, −7, 0, −4,2}, /* E */ {0, 2, −5, 3, 4, −5, 0, 1, −2, 0, 0, −3, −2, 1, _M, −1, 2,−1, 0, 0, 0, −2, −7, 0, −4, 3}, /* F */ {−4, −5, −4, −6, −5, 9, −5, −2,1, 0, −5, 2, 0, −4, _M, −5, −5, −4, −3, −3, 0, −1, 0, 0, 7, −5}, /* G */{1, 0, −3, 1, 0, −5, 5, −2, −3, 0, −2, −4, −3, 0, _M, −1, −1, −3, 1, 0,0, −1, −7, 0, −5, 0}, /* H */ {−1, 1, −3, 1, 1, −2, −2, 6, −2, 0, 0, −2,−2, 2, _M, 0, 3, 2, −1, −1, 0, −2, −3, 0, 0, 2}, /* I */ {−1, −2, −2,−2, −2, 1, −3, −2, 5, 0, −2, 2, 2, −2, _M, −2, −2, −2, −1, 0, 0, 4, −5,0, −1, −2}, /* J */ {0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, _M, 0, 0,0, 0, 0, 0, 0, 0, 0, 0, 0}, /* K */ {−1, 0, −5, 0, 0, −5, −2, 0, −2, 0,5, −3, 0, 1, _M, −1, 1, 3, 0, 0, 0, −2, −3, 0, −4, 0}, /* L */ {−2, −3,−6, −4, −3, 2, −4, −2, 2, 0, −3, 6, 4, −3, _M, −3, −2, −3, −3 , −1, 0,2, −2, 0, −1, −2} /* M */ {−1, −2, −5, −3, −2, 0, −3, −2, 2, 0, 0, 4, 6,−2, _M, −2, −1, 0, −2, −1, 0, 2, −4, 0, −2, −1}, /* N */ {0, 2, −4, 2,1, −4, 0, 2, −2, 0, 1, −3, −2, 2, _M, −1, 1, 0, 1, 0, 0, −2, −4, 0, −2,1}, /* O */ {_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,0,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,_M,} /* P */ {1, −1, −3, −1, −1, −5, −1,0, −2, 0, −1, −3, −2, −1, _M, 6, 0, 0, 1, 0, 0, −1, −6, 0, −5, 0}, /* Q*/ {0, 1, −5, 2, 2, −5, −1, 3, −2, 0, 1, −2, −1, 1, _M, 0, 4, 1, −1, −1,0, −2, −5, 0, −4, 3}, /* R */ {−2, 0, −4, −1, −1, −4, −3, 2, −2, 0, 3,−3, 0, 0, _M, 0, 1, 6, 0, −1, 0, −2, 2, 0, −4, 0}, /* S */ {1, 0, 0, 0,0, −3, 1, −1, −1, 0, 0, −3, −2, 1, _M, 1, −1, 0, 2, 1, 0, −1, −2, 0, −3,0}, /* T */ {1, 0, −2, 0, 0, −3, 0, −1, 0, 0, 0, −1, −1, 0, _M, 0, −1,−1, 1, 3, 0, 0, −5, 0, −3, 0}, /* U */ {0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0,0, 0, 0, _M, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0}, /* V */ {0, −2, −2, −2,−2, −1, −1, −2, 4, 0, −2, 2, 2, −2, _M, −1, −2, −2, −1, 0, 0, 4, −6, 0,−2, −2}, /* W */ {−6, −5, −8, −7, −7, 0, −7, −3, −5, 0, −3, −2, −4, −4,_M, −6, −5, 2, −2, −5, 0, −6, 17, 0, 0, −6}, /* X */ {0, 0, 0, 0, 0, 0,0, 0, 0, 0, 0, 0, 0, 0, _M, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0}, /* Y */{−3, −3, 0, −4, −4, 7, −5, 0, −1, 0, −4, −1, −2, −2, _M, −5, −4, −4, −3,−3, 0, −2, 0, 0, 10, −4}, /* Z */ {0, 1, −5, 2, 3, −5, 0, 2, −2, 0, 0,−2, −1, 1, _M, 0, 3, 0, 0, 0, 0, −2, −6, 0, −4, 4}, }; /*  */ #include<stdio.h> #include <ctype.h> #define MAXJMP  16 /* max jumps in a diag*/ #define MAXGAP  24 /* don't continue to penalize gaps larger thanthis */ #define JMPS 1024 /* max jmps in an path */ #define MX   4 /*save if there's at least MX-1 bases since last jmp */ #define DMAT   3/* value of matching bases */ #define DMIS   0 /* penalty for mismatchedbases */ #define DINS0   8 /* penalty for a gap */ #define DINS1   1 /*penalty per base */ #define PINS0   8 /* penalty for a gap */ #definePINS1   4 /* penalty per residue */ struct jmp { short n[MAXJMP]; /*size of jmp (neg for dely) */ unsigned short x[MAXJMP]; /* base no. ofjmp in seq x */ /* limits seq to 2 {circumflex over ( )}16 −1 */ };struct diag { int score; /* score at last jmp */ long offset; /* offsetof prev block */ short ijmp; /* current jmp index */ struct jmp jp; /*list of jmps */ }; struct path { int spc; /* number of leading spaces */short n[JMPS]; /* size of jmp (gap) */ int x[JMPS]; /* loc of jmp (lastelem before gap) */ }; char *ofile; /* output file name */ char*namex[2]; /* seq names: getseqs() */ char *prog; /* prog name for errmsgs */ char *seqx[2];   /* seqs: getseqs() */ int dmax; /* best diag:nw() */ int dmax0; /* final diag */ int dna; /* set if dna: main() */int endgaps; /* set if penalizing end gaps */ int gapx, gapy; /* totalgaps in seqs */ int len0, len1; /* seq lens */ int ngapx, ngapy; /*total size of gaps */ int smax; /* max score: nw() */ int *xbm; /*bitmap for matching */ long offset; /* current offset in jmp file */struct diag *dx; /* holds diagonals */ struct path pp[2]; /* holds pathfor seqs */ char *calloc(), *malloc(), *index(), *strcpy(); char*getseq(), *g_calloc(); /* Needleman-Wunsch alignment program  *  *usage: progs file1 file2  * where file1 and file2 are two dna or twoprotein sequences.  * The sequences can be in upper- or lower-case anmay contain ambiguity  * Any lines beginning with ‘;’, ‘>’ or ‘<’ areignored  * Max file length is 65535 (limited by unsigned short x in thejmp struct)  * A sequence with ⅓ or more of its elements ACGTU isassumed to be DNA  * Output is in the file “align.out”  *  * The programmay create a tmp file in /tmp to hold info about traceback.  * Originalversion developed under BSD 4.3 on a vax 8650  */ #include “nw.h”#include “day.h” static _dbval[26] = {1,14,2,13,0,0,4,11,0,0,12,0,3,15,0,0,0,5,6,8,8,7,9,0,10,0 }; static_pbval[26] = { 1, 2|(1< <(‘D’-‘A’))|(1< <(‘N’-‘A’)), 4, 8, 16, 32, 64,128, 256, 0×FFFFFFF, 1< <10, 1< <11, 1< <12, 1< <13, 1< <14, 1< <15, 1<<16, 1< <17, 1< <18, 1< <19, 1< <20, 1< <21, 1< <22, 1< <23, 1< <24, 1<<25|(1< <(‘E’-‘A’))|(1< <(‘Q’-‘A’)) }; main(ac, av) main int ac; char*av[]; { prog = av[0]; if (ac != 3) { fprintf(stderr, “usage: %s file1file2\n”, prog); fprintf(stderr, “where file1 and file2 are two dna ortwo protein sequences.\n”); fprintf(stderr, “The sequences can be inupper- or lower-case\n”); fprintf(stderr, “Any lines beginning with ‘;’or ‘<’ are ignored\n”); fprintf(stderr, “Output is in the file\“align.out\”\n”); exit(1); } namex[0] = av[1]; namex[1] = av[2];seqx[0] = getseq(namex[0], &len0); seqx[1] = getseq(namex[1], &len1);xbm = (dna)? _dbval : _pbval; endgaps = 0; /* 1 to penalize endgaps */ofile = “align.out”; /* output file */ nw(); /* fill in the matrix, getthe possible jmps */ readjmps(); /* get the actual jmps */ print(); /*print stats, alignment */ cleanup(0); /* unlink any tmp files */ } /* dothe alignment, return best score: main()  * dna: values in Fitch andSmith, PNAS, 80, 1382-1386, 1983  * pro: PAM 250 values  * When scoresare equal, we prefer mismatches to any gap, prefer  * a new gap toextending an ongoing gap, and prefer a gap in seqx  * to a gap in seq y. */ nw() nw { char *px, *py;   /* seqs and ptrs */ int *ndely, *dely; /*keep track of dely */ int ndelx, delx; /* keep track of delx */ int*tmp; /* for swapping row0, row1 */ int mis; /* score for each type */int ins0, ins1; /* insertion penalties */ register id; /* diagonal index*/ register ij; /* jmp index */ register *col0, *col1; /* score forcurr, last row */ register xx, yy; /* index into seqs */ dx = (structdiag *)g_calloc(“to get diags”, len0+len1+1, sizeof(struct diag)); ndely= (int *)g_calloc(“to get ndely”, len1+1, sizeof(int)); dely = (int*)g_calloc(“to get dely”, len1+1, sizeof(int)); col0 = (int*)g_calloc(“to get col0”, len1+1, sizeof(int)); col1 = (int*)g_calloc(“to get col1”, len1+1, sizeof(int)); ins0 = (dna)? DINS0 :PINS0; ins1 = (dna)? DINS1 : PINS1; smax = −10000; if (endgaps) { for(col0[0] = dely[0] = −ins0, yy = 1; yy < = len1; yy++) { col0[yy] =dely[yy] = col0[yy−1] − ins1; ndely[yy] = yy; } col0[0] = 0; /* WatermanBull Math Biol 84 */ } else for (yy = 1; yy < = len1; yy++) dely[yy] =−ins0; /* fill in match matrix  */ for (px = seqx[0], xx = 1; xx < =len0; px++, xx++) { /* initialize first entry in col  */ if (endgaps) {if (xx = = 1) col1[0] = delx = −(ins0+ins1); else col1[0] = delx =col0[0]−ins1; ndelx = xx; } else { col1[0] = 0; delx = −ins0; ndelx = 0;} ...nw for (py = seqx[1], yy = 1; yy < = len1; py++, yy++) { mis =col0[yy−1]; if (dna) mis += (xbm[*px−‘A’]&xbm[*py−‘A’])? DMAT : DMIS;else mis += _day[*px−‘A’][*py−‘A’]; /* update penalty for del in x seq; * favor new del over ongong del  * ignore MAXGAP if weighting endgaps */ if (endgaps || ndely[yy] < MAXGAP) { if (col0[yy] − ins0 > =dely[yy]) { dely[yy] = col0[yy] − (ins0+ins1); ndely[yy] = 1; } else {dely[yy] −= ins1; ndely[yy]++; } } else { if (col0[yy] − (ins0+ins1) > =dely[yy]) { dely[yy] = col0[yy] − (ins0+ins1); ndely[yy] = 1; } elsendely[yy]++; } /* update penalty for del in y seq;  * favor new del overongong del  */ if (endgaps || ndelx < MAXGAP) { if(col1[yy−1] − ins0 > =delx) { delx = col1[yy−1] − (ins0+ins1); ndelx = 1; } else { delx −=ins1; ndelx++; } } else { if (col1[yy−1] − (ins0+ins1) >= delx) { delx =col1[yy−1] − (ins0+ins1); ndelx = 1; } else ndelx++; } /* pick themaximum score; we're favoring  * mis over any del and delx over dely  */...nw id = xx − yy + len1 − 1; if (mis > = delx && mis > = dely[yy])col1[yy] = mis; else if (delx > = dely[yy]) { col1[yy] = delx; ij =dx[id].ijmp; if (dx[id].jp.n[0] && (!dna || (ndelx > = MAXJMP && xx >dx[id].jp.x[ij]+MX) || mis > dx[id].score+DINS0)) { dx[id].ijmp++; if(++ij > = MAXJMP) { writejmps(id); ij = dx[id].ijmp = 0; dx[id].offset =offset; offset += sizeof(struct jmp) + sizeof(offset); } }dx[id].jp.n[ij] = ndelx; dx[id].jp.x[ij] = xx; dx[id].score = delx; }else { col1[yy] = dely[yy]; ij = dx[id].ijmp; if (dx[id].jp.n[0] &&(!dna || (ndely[yy] > = MAXJMP && xx > dx[id].jp.x[ij]+MX) || mis >dx[id].score+DINS0)) { dx[id].ijmp++; if (++ij > = MAXJMP) {writejmps(id); ij = dx[id].ijmp = 0; dx[id].offset = offset; offset +=sizeof(struct jmp) + sizeof(offset); } } dx[id].jp.n[ij] = −ndely[yy];dx[id].jp.x[ij] = xx; dx[id].score = dely[yy]; } if (xx = = len0 && yy <len1) { /* last col  */ if (endgaps) col1[yy] −= ins0+ins1*(len1−yy);if(col1[yy] > smax) { smax = col1[yy]; dmax = id; } } } if (endgaps &&xx < len0) col1[yy−1] −= ins0+ins1*(len0−xx); if (col1[yy−1] > smax) {smax = col1[yy−1]; dmax = id; } tmp = col0; col0 = col1; col1 = tmp; }(void) free((char *)ndely); (void) free((char *)dely); (void) free((char*)col0); (void) free((char *)col1); } /*  *  * print() -- only routinevisible outside this module  *  * static:  * getmat() -- trace back bestpath, count matches: print()  * pr_align() -- print alignment ofdescribed in array p[]: print()  * dumpblock() -- dump a block of lineswith numbers, stars: pr_align()  * nums() -- put out a number line:dumpblock()  * putline() -- put out a line (name, [num], seq, [num]):dumpblock()  * stars() - -put a line of stars: dumpblock()  *stripname() -- strip any path and prefix from a seqname  */ #include“nw.h” #define SPC  3 #define P_LINE 256 /* maximum output line */#define P_SPC  3 /* space between name or num and seq */ extern_day[26][26]; int olen; /* set output line length */ FILE *fx; /* outputfile */ print() print { int lx, ly, firstgap, lastgap;  /* overlap */ if((fx = fopen(ofile, “w”)) = = 0) { fprintf(stderr, “%s: can't write%s\n”, prog, ofile); cleanup(1); } fprintf(fx, “<first sequence: %s(length = %d)\n”, namex[0], len0); fprintf(fx, “<second sequence: %s(length = %d)\n”, namex[1], len1); olen = 60; lx = len0; ly = len1;firstgap = lastgap = 0; if (dmax < len1 − 1) { /* leading gap in x */pp[0].spc = firstgap = len1 − dmax − 1; ly −= pp[0].spc; } else if(dmax > len1 − 1) { /* leading gap in y */ pp[1].spc = firstgap = dmax −(len1 − 1); lx −= pp[1].spc; } if (dmax0 < len0 − 1) { /* trailing gapin x */ lastgap = len0 − dmax0 −1; lx −= lastgap; } else if (dmax0 >len0 − 1) { /* trailing gap in y */ lastgap = dmax0 − (len0 − 1); ly −=lastgap; } getmat(lx, ly, firstgap, lastgap); pr_align(); } /*  * traceback the best path, count matches  */ static getmat(lx, ly, firstgap,lastgap) getmat int lx, ly; /* “core” (minus endgaps) */ int firstgap,lastgap; /* leading trailing overlap */ { int nm, i0, i1, siz0, siz1;char outx[32]; double pct; register n0, n1; register char *p0, *p1; /*get total matches, score  */ i0 = i1 = siz0 = siz1 = 0; p0 = seqx[0] +pp[1].spc; p1 = seqx[1] + pp[0].spc; n0 = pp[1].spc + 1; n1 =pp[0].spc + 1; nm = 0; while ( *p0 && *p1 ) { if (siz0) { p1++; n1++;siz0−−; } else if (siz1) { p0++; n0++; siz1−−; } else { if(xbm[*p0−‘A’]&xbm[*p1−‘A’]) nm++; if (n0++ = = pp[0].x[i0]) siz0 =pp[0].n[i0++]; if (n1++ = = pp[1].x[i1]) siz1 = pp[1].n[i1++]; p0++;p1++; } } /* pct homology:  * if penalizing endgaps, base is the shorterseq  * else, knock off overhangs and take shorter core  */ if (endgaps)lx = (len0 < len1)? len0 : len1; else lx = (lx < ly)? lx : ly; pct =100.*(double)nm/(double)lx; fprintf(fx, “\n”); fprintf(fx, “<%d match%sin an overlap of %d: %.2f percent similarity\n”, nm, (nm = = 1)? “” :“es”, lx, pct); fprintf(fx, “<gaps in first sequence: %d”, gapx);...getmat if (gapx) { (void) sprintf(outx, “ (%d %s%s)”, ngapx, (dna)?“base”:“residue”, (ngapx = = 1)? “”:“s”); fprintf(fx, “%s”, outx);fprintf(fx, “, gaps in second sequence: %d”, gapy); if (gapy) { (void)sprintf(outx, “ (%d %s%s)”, ngapy, (dna)? “base”:“residue”, (ngapy = =1)? “”:“s”); fprintf(fx,“%s”, outx); } if (dna) fprintf(fx, “\n<score:%d (match = %d, mismatch = %d, gap penalty = %d + %d per base)\n”, smax,DMAT, DMIS, DINS0, DINS1); else fprintf(fx, “\n<score: %d (Dayhoff PAM250 matrix, gap penalty = %d + %d per residue)\n”, smax, PINS0, PINS1);if (endgaps) fprintf(fx, “<endgaps penalized. left endgap: %d %s%s,right endgap: %d %s%s\n”, firstgap, (dna)? “base” : “residue”, (firstgap= = 1)? “” : “s”, lastgap, (dna)? “base” : “residue”, (lastgap = = 1)?“” : “s”); else fprintf(fx, “<endgaps not penalized\n”); } static nm; /*matches in core -- for checking */ static lmax; /* lengths of strippedfile names */ static ij[2]; /* jmp index for a path */ static nc[2]; /*number at start of current line */ static ni[2]; /* current elem number-- for gapping */ static siz[2]; static char *ps[2]; /* ptr to currentelement */ static char *po[2]; /* ptr to next output char slot */ staticchar out[2][P_LINE]; /* output line */ static char star[P_LINE]; /* setby stars() */ /*  * print alignment of described in struct path pp[]  */static pr_align() pr_align { int nn; /* char count */ int more; registeri; for (i = 0, lmax = 0; i < 2; i++) { nn = stripname(namex[i]); if(nn > lmax) lmax = nn; nc[i] = 1; ni[i] = 1; siz[i] = ij[i] = 0; ps[i] =seqx[i]; po[i] = out[i]; } for (nn = nm = 0, more = 1; more;) {...pr_align for (i = more = 0; i < 2; i++) { /*  * do we have more ofthis sequence?  */ if (!*ps[i]) continue; more++; if (pp[i].spc) { /*leading space */ *po[i]++ = ‘ ’; pp[i].spc−−; } else if (siz[i]) { /* ina gap */ *po[i]++ = ‘−’; siz[i]−−; } else { /* we're putting a seqelement  */ *po[i] = *ps[i]; if (islower(*ps[i])) *ps[i] =toupper(*ps[i]); po[i]++; ps[i]++; /*  * are we at next gap for thisseq?  */ if (ni[i] = = pp[i].x[ij[i]]) { /*  * we need to merge all gaps * at this location  */ siz[i] = pp[i].n[ij[i]++]; while (ni[i] = =pp[i].x[ij[i]]) siz[i] += pp[i].n[ij[i]++]; } ni[i]++; } } if (++nn = =olen || !more && nn) { dumpblock(); for (i = 0; i < 2; i++) po[i] =out[i]; nn = 0; } } } /*  * dump a block of lines, including numbers,stars: pr_align()  */ static dumpblock() dumpblock { register i; for (i= 0; i < 2; i++) *po[i]−− = ‘\0’; ...dumpblock (void) putc(‘\n’, fx);for (i = 0; i < 2; i++) { if (*out[i] && (*out[i] != ‘ ’ || *(po[i]) !=‘ ’)) { if (i = = 0) nums(i); if (i = = 0 && *out[1]) stars();putline(i); if (i = = 0 && *out[1]) fprintf(fx, star); if (i = = 1)nums(i); } } } /*  * put out a number line: dumpblock()  */ staticnums(ix) nums int  ix; /* index in out[] holding seq line */ { charnline[P_LINE]; register i, j; register char *pn, *px, *py; for (pn =nline, i = 0; i < lmax+P_SPC; i++, pn++) *pn = ‘ ’; for (i = nc[ix], py= out[ix]; *py; py++, pn++) { if (*py = = ‘ ’ || *py = = ‘−’) *pn = ‘ ’;else { if (i%10 = = 0 || (i = = 1 && nc[ix] != 1)) { j = (i < 0)? −i :i; for (px = pn; j; j /= 10, px−−) *px = j%10 + ‘0’; if (i < 0) *px =‘−’; } else *pn = ‘ ’; i++; } } *pn = ‘\0’; nc[ix] = i; for (pn = nline;*pn; pn++) (void) putc(*pn, fx); (void) putc(‘\n’, fx); } /*  * put outa line (name, [num], seq, [num]): dumpblock()  */ static putline(ix)putline int   ix; { ...putline int i; register char *px; for (px =namex[ix], i = 0; *px && *px != ‘:’; px++, i++) (void) putc(*px, fx);for (; i < lmax+P_SPC; i++) (void) putc(‘ ’, fx); /* these count from 1: * ni[] is current element (from 1)  * nc[] is number at start ofcurrent line  */ for (px = out[ix]; *px; px++) (void) putc(*px&0x7F,fx); (void) putc(‘\n’, fx); } /*  * put a line of stars (seqs always inout[0], out[1]): dumpblock()  */ static stars() stars { int i; registerchar *p0, *p1, cx, *px; if (!*out[0] || (*out[0] = = ‘ ’ && *(po[0]) = =‘ ’) || !*out[1] || (*out[1] = = ‘ ’ && *(po[1]) = = ‘ ’)) return; px =star; for (i = lmax+P_SPC; i; i−−) *px++ = ‘ ’; for (p0 = out[0], p1 =out[1]; *p0 && *p1; p0++, p1++) { if (isalpha(*p0) && isalpha(*p1)) { if(xbm[*p0−‘A’]&xbm[*p1−‘A’]) { cx = ‘*’; nm++; } else if (!dna &&_day[*p0−‘A’][*p1−‘A’] > 0) cx = ‘.’; else cx = ‘ ’; } else cx = ‘ ’;*px++ = cx; } *px++ = ‘\n’; *px = ‘\0’; } /*  * strip path or prefixfrom pn, return len: pr_align()  */ static stripname(pn) stripname char*pn; /* file name (may be path) */ { register char *px, *py; py = 0; for(px = pn; *px; px++) if (*px = = ‘/’) py = px + 1; if (py) (void)strcpy(pn, py); return(strlen(pn)); } /*  * cleanup() -- cleanup any tmpfile  * getseq() -- read in seq, set dna, len, maxlen  * g_calloc() --calloc() with error checkin  * readjmps() -- get the good jmps, from tmpfile if necessary  * writejmps() -- write a filled array of jmps to atmp file: nw()  */ #include “nw.h” #include <sys/file.h> char *jname =“/tmp/homgXXXXXX”; /* tmp file for jmps */ FILE *fj; int cleanup(); /*cleanup tmp file */ long lseek(); /*  * remove any tmp file if we blow */ cleanup(i) cleanup int i; { if (fj) (void) unlink(jname); exit(i); }/*  * read, return ptr to seq, set dna, len, maxlen  * skip linesstarting with ‘;’, ‘<’, or ‘>’  * seq in upper or lower case  */ char  *getseq(file, len) getseq char *file; /* file name */ int *len; /* seqlen */ { char line[1024], *pseq; register char *px, *py; int natgc,tlen; FILE *fp; if ((fp = fopen(file, “r”)) = = 0) { fprintf(stderr,“%s: can't read %s\n”, prog, file); exit(1); } tlen = natgc = 0; while(fgets(line, 1024, fp)) { if (*line = = ‘;’ || *line = = ‘<’ || *line= = ‘>’) continue; for (px = line; *px != ‘\n’; px++) if (isupper(*px)|| islower(*px)) tlen++; } if ((pseq = malloc((unsigned)(tlen+6))) = =0) { fprintf(stderr, “%s: malloc() failed to get %d bytes for %s\n”,prog, tlen+6, file); exit(1); } pseq[0] = pseq[1] = pseq[2] = pseq[3] =‘\0’; ...getseq py = pseq + 4; *len = tlen; rewind(fp); while(fgets(line, 1024, fp)) { if (*line = = ‘;’ || *line = = ‘<’ || *line= = ‘>’) continue; for (px = line; *px != ‘\n’; px++) { if(isupper(*px)) *py++ = *px; else if (islower(*px)) *py++ = toupper(*px);if (index(“ATGCU”, *(py−1))) natgc++; } } *py++ = ‘\0’; *py = ‘\0’;(void) fclose(fp); dna = natgc > (tlen/3); return(pseq+4); } char *g_calloc(msg, nx, sz) g_calloc char *msg; /* program, calling routine */int nx, sz; /* number and size of elements */ { char *px, *calloc(); if((px = calloc((unsigned)nx, (unsigned)sz)) = = 0) { if (*msg) {fprintf(stderr, “%s: g_calloc() failed %s (n= %d, sz= %d)\n”, prog, msg,nx, sz); exit(1); } } return(px); } /*  * get final jmps from dx[] ortmp file, set pp[], reset dmax: main()  */ readjmps() readjmps { int fd= −1; int siz, i0, i1; register i, j, xx; if (fj) { (void) fclose(fj);if ((fd = open(jname, O_RDONLY, 0)) < 0) { fprintf(stderr, “%s: can'topen() %s\n”, prog, jname); cleanup(1); } } for (i = i0 = i1 = 0, dmax0= dmax, xx = len0; ; i++) { while (1) { for (j = dx[dmax].ijmp; j > = 0&& dx[dmax].jp.x[j] > = xx; j−−) ; ...readjmps if (j < 0 &&dx[dmax].offset && fj) { (void) lseek(fd, dx[dmax].offset, 0); (void)read(fd, (char *)&dx[dmax].jp, sizeof(struct jmp)); (void) read(fd,(char *)&dx[dmax].offset, sizeof(dx[dmax].offset)); dx[dmax].ijmp =MAXJMP−1; } else break; } if (i >= JMPS) { fprintf(stderr, “%s: too manygaps in alignment\n”, prog); cleanup(1); } if (j > = 0) { siz =dx[dmax].jp.n[j]; xx = dx[dmax].jp.x[j]; dmax += siz; if (siz < 0) { /*gap in second seq */ pp[1].n[i1] = −siz; xx += siz; /* id = xx − yy +len1 − 1  */ pp[1].x[i1] = xx − dmax + len1 − 1; gapy++; ngapy −= siz;/* ignore MAXGAP when doing endgaps */ siz = (−siz < MAXGAP || endgaps)?−siz : MAXGAP; i1++; } else if (siz > 0) { /* gap in first seq */pp[0].n[i0] = siz; pp[0].x[i0] = xx; gapx++; ngapx += siz; /* ignoreMAXGAP when doing endgaps */ siz = (siz < MAXGAP || endgaps)? siz :MAXGAP; i0++; } } else break; } /* reverse the order of jmps  */ for (j= 0, i0−−; j < i0; j++, i0−−) { i = pp[0].n[j]; pp[0].n[j] =pp[0].n[i0]; pp[0].n[i0] = i; i = pp[0].x[j]; pp[0].x[j] = pp[0].x[i0];pp[0].x[i0] = i; } for (j = 0, i1−−; j < i1; j++, i1−−) { i =pp[1].n[j]; pp[1].n[j] = pp[1].n[i1]; pp[1].n[i1] = i; i = pp[1].x[j];pp[1].x[j] = pp[1].x[i1]; pp[1].x[i1] = i; } if (fd > = 0) (void)close(fd); if (fj) { (void) unlink(jname); fj = 0; offset = 0; } } /*  *write a filled jmp struct offset of the prev one (if any): nw()  */writejmps(ix) writejmps int ix; { char *mktemp(); if (!fj) { if(mktemp(jname) < 0) { fprintf(stderr, “%s: can't mktemp() %s\n”, prog,jname); cleanup(1); } if ((fj = fopen(jname, “w”)) = = 0) {fprintf(stderr, “%s: can't write %s\n”, prog, jname); exit(1); } }(void) fwrite((char *)&dx[ix].jp, sizeof(struct jmp), 1, fj); (void)fwrite((char *)&dx[ix].offset, sizeof(dx[ix].offset), 1, fj); }

[1103] TABLE 2 PRO XXXXXXXXXXXXXXX (Length = 15 amino acids) ComparisonXXXXXYYYYYYY (Length = 12 amino acids) Protein % amino acid sequenceidentity = (the number of identically matching amino acid residuesbetween the two polypeptide sequences as determined by ALIGN-2) dividedby (the total number of amino acid residues of the PRO polypeptide) = 5divided by 15 = 33.3%

[1104] TABLE 3 PRO XXXXXXXXXX (Length = 10 amino acids) ComparisonXXXXXYYYYYYZZYZ (Length = 15 amino acids) Protein % amino acid sequenceidentity = (the number of identically matching amino acid residuesbetween the two polypeptide sequences as determined by ALIGN-2) dividedby (the total number of amino acid residues of the PRO polypeptide) = 5divided by 10 = 50%

[1105] TABLE 4 PRO-DNA NNNNNNNNNNNNNN (Length = 14 nucleotides)Comparison NNNNNNLLLLLLLLLL (Length = 16 nucleotides) DNA % nucleic acidsequence identity = (the number of identically matching nucleotidesbetween the two nucleic acid sequences as determined by ALIGN-2) dividedby (the total number of nucleotides of the PRO-DNA nucleic acidsequence) = 6 divided by 14 = 42.9%

[1106] TABLE 5 PRO-DNA NNNNNNNNNNNN (Length = 12 nucleotides) ComparisonDNA NNNNLLLVV (Length = 9 nucleotides) % nucleic acid sequence identity= (the number of identically matching nucleotides between the twonucleic acid sequences as determined by ALIGN-2) divided by (the totalnumber of nucleotides of the PRO-DNA nucleic acid sequence) = 4 dividedby 12 = 33.3%

[1107] II. Compositions and Methods of the Invention

[1108] A. Full-Length PRO Polypeptides

[1109] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO polypeptides. In particular, cDNAs encoding variousPRO polypeptides have been identified and isolated, as disclosed infurther detail in the Examples below. It is noted that proteins producedin separate expression rounds may be given different PRO numbers but theUNQ number is unique for any given DNA and the encoded protein, and willnot be changed. However, for sake of simplicity, in the presentspecification the protein encoded by the full length native nucleic acidmolecules disclosed herein as well as all further native homologues andvariants included in the foregoing definition of PRO, will be referredto as “PRO/number”, regardless of their origin or mode of preparation.

[1110] As disclosed in the Examples below, various cDNA clones have beendeposited with the ATCC. The actual nucleotide sequences of those clonescan readily be determined by the skilled artisan by sequencing of thedeposited clone using routine methods in the art. The predicted aminoacid sequence can be determined from the nucleotide sequence usingroutine skill. For the PRO polypeptides and encoding nucleic acidsdescribed herein, Applicants have identified what is believed to be thereading frame best identifiable with the sequence information availableat the time.

[1111] 1. Full-Length PRO213 Polypeptides

[1112] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO213. In particular, Applicants have identified andisolated cDNA encoding a PRO213 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that a portion of the PRO213polypeptide has significant homology with the human growtharrest-specific 6 (gas6) protein. Accordingly, it is presently believedthat PRO213 polypeptide disclosed in the present application may havethe same or simular activity as does the gas6 protein.

[1113] 2. Full-Length PRO274 Polypeptides

[1114] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO274. In particular, Applicants have identified andisolated cDNA encoding a PRO274 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO274polypeptide have significant homology with the 7 transmembrane segmentreceptor proteins and Fn54 protein. Accordingly, it is presentlybelieved that PRO274 polypeptide disclosed in the present application isa newly identified member of the 7 transmembrane segment receptorprotein and/or Fn54 protein family.

[1115] 3. Full-Length PRO300 Polypeptides

[1116] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO300. In particular, Applicants have identified andisolated cDNA encoding a PRO300 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO300polypeptide have significant homology with the human Diff 33 protein.Accordingly, it is presently believed that PRO300 polypeptide disclosedin the present application is a newly identified member of the Diff 33family.

[1117] 4. Full-Length PRO284 Polypeptides

[1118] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO284. In particular, Applicants have identified andisolated cDNA encoding a PRO284 polypeptide, as disclosed in frtherdetail in the Examples below. To Applicants present knowledge, theUNQ247 (DNA23318-1211) nucleotide sequence encodes a novel factor; usingBLAST and FastA sequence alignment computer programs, no sequenceidentities to any known proteins were revealed.

[1119] 5. Full-Length PRO296 Polypeptides

[1120] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO296. In particular, Applicants have identified andisolated cDNA encoding a PRO296 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO296 polypeptide hassignificant similarity to the sarcoma-amplified SAS protein.Accordingly, it is presently believed that PRO296 polypeptide disclosedin the present application is a newly identified SAS protein homolog. 6.Full-Length PRO329 Polypeptides

[1121] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO329. In particular, Applicants have identified andisolated cDNA encoding a PRO329 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO329 polypeptide hassignificant similarity to a high affinity inmmunoglobulin F. receptor.Accordingly, it is presently believed that PRO329 polypeptide disclosedin the present application is a newly identified F. receptor homolog.

[1122] 7. Full length PRO362 Polypeptides

[1123] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO362. In particular, Applicants have identified andisolated cDNA encoding a PRO362 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO362 polypeptide hassignificant similarity to the A33 antigen protein as well as the HCARprotein and the NrCAM related cell adhesion molecule. Accordingly, it ispresently believed that PRO362 polypeptide disclosed in the presentapplication is a newly A33 antigen and HCAR protein homolog.

[1124] 8. Full-Length PRO363 Polypeptides

[1125] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO363. In particular, Applicants have identified andisolated cDNA encoding a PRO363 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO363 polypeptide hassignificant similarity to the cell surface protein HCAR. Accordingly, itis presently believed that PRO363 polypeptide disclosed in the presentapplication is a newly HCAR homolog.

[1126] 9. Full-Length PRO868 Polypeptides

[1127] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO868. In particular, Applicants have identified andisolated cDNA encoding a PRO868 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO868 polypeptide hassignificant similarity to the tumor necrosis factor receptor.Accordingly, it is presently believed that PRO868 polypeptide disclosedin the present application is a newly identified member of the tumornecrosis factor receptor family of proteins.

[1128] 10. Full-Length PRO382 Polypeptides

[1129] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO382. In particular, Applicants have identified andisolated cDNA encoding a PRO382 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence aligmnentcomputer programs, Applicants found that the native PRO382 polypeptideshares significant homology with various serine protease proteins.Applicants have also found that the DNA encoding the PRO382. polypeptideshares significant homology with nucleic acid encoding various serineprotease proteins. Accordingly, it is presently believed that PRO382polypeptide disclosed in the present application is a newly identifiedserine protease homolog.

[1130] 11. Full-Length PRO545 Polypeptides

[1131] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO545. In particular, Applicants have identified andisolated cDNA encoding a PRO545 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO545polypeptide have significant homology with the sequences identifieddesignated as; human metalloproteinase (“P_WO1825”), mouse meltrin alpha(“S60257”), metalloprotease-disintegrin meltrin-alpha (“GEN13695 ”),ADAM 13—Xenopus laevis (“XLU66003_(—)1”), mouse meltrin beta (“S60258”),rabbit metalloprotease-disintegrin meltrin-beta, (“GEN13696”), humanmeltrin S (“AF023477_(—)1”), human meltrin precursor (“AF023476_(—)1”),human ADAM 21 (AF029900_(—)1”), and human ADAM 20 (“AF0298991_(—)1),thereby indicating that PRO545 may be a novel meltrin protein.Accordingly, it is presently believed that the PRO545 polypeptidedisclosed in the present application is a newly identified member of themeltrin family and possesses the cellular adhesiveness typical of themeltrin proteins which comprise both metalloprotease and disintegrindomains.

[1132] 12. Full-Length PRO617 Polypeptides

[1133] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO617. In particular, Applicants have identified andisolated cDNA encoding a PRO617 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO617 polypeptide sharessignificant homology with the CD24 protein. Applicants have also foundthat the DNA encoding the PRO617 polypeptide has significant homologywith DNA encoding the CD24 protein. Accordingly, it is presentlybelieved that PRO617 polypeptide disclosed in the present application isa newly identified CD24 homolog.

[1134] 13. Full-Length PRO700 Polypeptides

[1135] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO700. In particular, Applicants have identified andisolated cDNA encoding a PRO700 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO700 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO700polypeptide possess significant sequence similarity to various proteindisulfide isomerases. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant sequencesimilarity between the PRO700 amino acid sequence and the followingDayhoff sequences; polypeptide with protein disulfide isomeraseactivity, designated as (“P_P80664”), human PDI, designated as(“P_R51696”), human PDI, designated as (P_R5297”), probable proteindisulfide isomerase er-60 precursor, designated as (“ER60_SCMA”),protein disulfide isomerase precursor—Drosophila melanogaster,designated as (“PDI_DROME”), protein disulfide-isomeraseprecursor—Nicotiana tabaccum, designated as (“NTFDIGENE_(—)1,), proteindisulfide isomerase—Onchocerca volvulus, designated as(“OVU12440_(—)1”), human probable protein disulfide isomerase p5precursor, designated as (ERP5_HUMAN”), human protein disulfideisomerase-related protein 5, (“HSU79278_(—)1”), and protein disulfideisomerase precursor/prolyl 4 hydroxy, (“PDI_HUMAN”), thereby indicatingthat PRO700 may be a novel protein disulfide isomerase. Accordingly, itis presently believed that PRO700 polypeptide disclosed in the presentapplication is a newly identified member of the protein disulfideisomerase family and possesses the ability to catalyze the formation ofdisulfide bonds typical of the protein disulfide isomerase family.

[1136] 14. Full-Length PRO702 Polypeptides

[1137] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO702. In particular, Applicants have identified andisolated cDNA encoding a PRO702 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO702 polypeptide hassignificant similarity to the conglutinin protein. Accordingly, it ispresently believed that PRO702 polypeptide disclosed in the presentapplication is a newly identified conglutinin homolog.

[1138] 15. Full-Length PRO703 Polypeptides

[1139] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO703. In particular, Applicants have identified andisolated cDNA encoding a PRO703 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amnino acid sequence ofthe full-length PRO703 polypeptide using BLAST and FastA sequencealignment computer programs, suggests that various portions of thePRO703 polypeptide possess significant sequence similarity to the VLCASprotein, thereby indicating that PRO703 may be a novel VLCAS protein.More specifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant sequence similarity between thePRO703 amino acid sequence and the following Dayhoff sequences, humanmRNA for very-long-chain acyl-CoA, (“D88308”), rat MnRNA forvery-long-chain acyl-CoA synthetase, (“D85100”), Mus musculs fatty acidtransport protein, (“MMU15976”), human very-long-chain acyl-CoAsynthetase, (“D88308_(—)1”), Mus musculus very-long-chain acyl-CoAsynthetase, (“AP033031_(—)1”), very-long-chain acyl-CoAsynthetase—Rauus, (“D85100_(—)1”), rat long-chain fatty acid transportprotein, (“FATP_RAT”), mouse long-chain fatty acid transport protein,(“FATP_MOUSE”), probable long-chain fatty acid transport protein,(“FAT1_YEAST”), and fatty acid transporter protein, (“CHY15839_(—)2”),thereby indicating that PRO703 may be a novel VLCAS. Accordingly, it ispresently believed that PRO703 polypeptide disclosed in the presentapplication is a newly identified member of the VLCAS family andpossesses the ability to facilitate the cellular transport of long andvery long chain fatty acids typical of the VLCAS family.

[1140] 16. Full-Length PRO705 Polypeptides

[1141] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO705. In particular, Applicants have identified andisolated cDNA encoding a PRO705 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO705 polypeptide hassignificant similarity to the K-glypican protein. Accordingly, it ispresently believed that PRO705 polypeptide disclosed in the presentapplication is a newly identified member of the glypican family ofproteoglycan proteins.

[1142] 17. Full-Length PRO708 Polypeptides

[1143] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO708. In particular, Applicants have identified andisolated cDNA encoding a PRO708 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO708 polypeptide hassignificant homology with the aryl sulfatase proteins. Applicants havealso found that the DNA encoding the PRO708 polypeptide has significanthomology with DNA encoding the aryl sulfatase proteins. Accordingly, itis presently believed that PRO708 polypeptide disclosed in the presentapplication is a newly identified aryl sulfatase homolog.

[1144] 18. Full-Length PRO320 Polypeptides

[1145] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO320. In particular, Applicants have identified andisolated cDNA encoding a PRO320 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO320 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO320polypeptide have significant homology to the fibulin protein. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant homology between the PRO320 aminoacid sequence and the following Dayhoff sequences, human fibulin-2precursor, designated “FBL2_HUMAN”, human fibulin-I isoform a precursor,designated “FBLA_HUMAN”, ZK783.1—Caenorhabditis elegans, designated“CELZK783_(—)1”, human-notcb2, designated “HSU77493_(—)1”, Nel proteinprecursor—rattus norvegicus, designated “NEL_RAT”, Mus musculus cellsurface protein, designated “D32210_(—)1”, mouse (fragment) Notch Bprotein, designated “A49175”, C50H2.3a—Caenorhabditis elegans,designated “CEC50H2_(—)3”, MEC-9L—Caenorhabditis elegans, designated“CEU33933_(—)1”, and Mus musculus notch 4, designated “10MMMHC29N7_(—)2”, thereby indicating that PRO320 may be a novel fibulinor fibulin-like protein. Accordingly, it is presently believed thatPRO320 polypeptide disclosed in the present application is a newlyidentified member of the fibulin family and possesses biologicalactivity typical of the fibulin family.

[1146] 19. Full-Length PRO324 Polypeptides

[1147] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO324. In particular, Applicants have identified andisolated cDNA encoding a PRO324 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO324 polypeptide hassignificant similarity to oxidoreductases. Accordingly, it is presentlybelieved that PRO324 polypeptide disclosed in the present application isa newly identified oxidoreductase homolog.

[1148] 20. Full-Length PRO351 Polypeptides

[1149] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO351. In particular, Applicants have identified andisolated cDNA encoding a PRO351 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO351 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO351polypeptide possess significant sequence similarity to the prostasinprotein, thereby indicating that PRO351 may be a novel prostasinprotein. More specifically, an analysis of the Dayhoff database (version35.45 SwissProt 35) evidenced significant sequence similarity betweenthe PRO351 amino acid sequence and the following Dayhoff sequences,“AC003965_(—)1”, “CELC07G1_(—)7”, “GEN12917”, “HEPS_HUMAN”, “GEN14584”,“MCT6_MOUSE”, “HSU75329_(—)1”, “PLMN_ERIEU”, “TRYB_HUMAN”, and“P_W22987”. Accordingly, it is presently believed that PRO351polypeptide disclosed in the present application is a newly identifiedmember of the prostasin family and possesses properties and activitiestypical of the prostasin family.

[1150] 21. Full-Length PRO352 Polypeptides

[1151] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO352. In particular, Applicants have identified andisolated cDNA encoding a PRO352 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO352 polypeptide hassignificant similarity to the butyrophilin protein. Accordingly, it ispresently believed that PRO352 polypeptide disclosed in the presentapplication is a newly identified butyrophilin homolog.

[1152] 22. Full-Length PRO381 Polypeptides

[1153] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO381. In particular, Applicants have identified andisolated cDNA encoding a PRO381 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO381 polypeptide hassignificant similarity to immunophilin proteins. Accordingly, it ispresently believed that PRO381 polypeptide disclosed in the presentapplication is a newly identified FKBP immunophilin homolog.

[1154] 23. Full-Length PRO386 Polypeptides

[1155] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO386. In particular, Applicants have identified andisolated cDNA encoding a PRO386 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO386polypeptide hassignificant similarity to the beta-2 subunit of a sodium channelprotein. Accordingly, it is presently believed that PRO386 polypeptidedisclosed in the present application is homolog of a beta-2 subunit of asodium channel expressed in mammalia cells.

[1156] 24. Full-Length PRO540 Polypeptides

[1157] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO540. In particular, Applicants have identified andisolated cDNA encoding a PRO540 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO540 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO540polypeptide possess significant sequence similarity to the LCAT protein,thereby indicating that PRO540 may be a novel LCAT protein. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant sequence similarity between thePRO540 amino acid sequence and the following Dayhoff sequences,phosphatidylcholine-sterol acyltransferase, designated “LCAT_HUMAN”,hypothetical 75.4 kd protein, designated “YN84_YEAST”, Bacilluslicheniforms esterase, designated “BLU35855_(—)1”, macrotetrolideresistance protein—Streptomyces, designated “JH0655”, T-cell receptordelta chain precursor, designated “C30583”, Rhesus kringle 2, designated“P_W07551”, RAGE-1 ORF5, designated “HSU46191_(—)3”, human Ig kappachain VKIII-JK3, designated “HSU07466_(—)1”, and Alstroemeria inodorareverse transcriptase, designated “AL1223606_(—)1”. Accordingly, it ispresently believed that PRO540 polypeptide disclosed in the presentapplication is a newly identified member of the LCAT protein family andpossesses lipid transport capability typical of the LCAT family.

[1158] 25. Full-Length PRO615 Polypeptides

[1159] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO615. In particular, Applicants have identified andisolated cDNA encoding a PRO615 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO615 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO615polypeptide possess significant sequence similarity to the humansynaptogyrin protein, thereby indicating that PRO615 may be a novelsynaptogyrin protein. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant sequencesimilarity between the PRO615 amino acid sequence and the followingDayhoff sequences, “AF039085_(—)1”, “RNU39549_(—)1”, “CELT08A9_(—)8”,“FSU62028_(—)1”, “S73645”, “Y348_MYCPN”, “AC000103_(—)5”, “ ”,“RT12_LEITA”, and “EBVLMP218_(—)1”. Accordingly, it is presentlybelieved that PRO615 polypeptide disclosed in the present application isa newly identified member of the synaptogyrin family and possessesactivity and properties typical of the synaptogyrin family.

[1160] 26. Full-Length PRO618 Polypeptides

[1161] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO618. In particular, Applicants have identified andisolated cDNA encoding a PRO618 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO618 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO618polypeptide possess significant sequence similarity to theenteropeptidase protein, thereby indicating that PRO618 may be a novelenteropeptidase. More specifically, an analysis of the Dayhoff database(version 35.45 SwissProt 35) evidenced significant sequence similaritybetween the PRO618 amino acid sequence and the following Dayhoffsequences, “P_W22987”, “KAL_HUMAN”, “AC00395_(—)1”, “GEN12917”,“ENTK_HUMAN”, “FA11_HUMAN”, “HSU75329_(—)1”, “P_W22986”, and“PLMN_HORSE”. Accordingly, it is presently believed that PRO618polypeptide disclosed in the present application is a newly identifiedmember of the enteropeptidase family and possesses catalytic activitytypical of the enteropeptidase family.

[1162] 27. Full-Length PRO719 Polypeptides

[1163] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO719. In particular, Applicants have identified andisolated cDNA encoding a PRO719 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO719 polypeptide hassignificant similarity to the lipoprotein lipase H protein. Accordingly,it is presently believed that PRO719 polypeptide disclosed in thepresent application is a newly identified lipoprotein lipase H homolog.

[1164] 28. Full-Length PRO724 Polypeptides

[1165] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO724. In particular, Applicants have identified andisolated cDNA encoding a PRO724 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO724 polypeptide hassignificant similarity to the human low density lipoprotein (LDL)receptor protein. Accordingly, it is presently believed that PRO724polypeptide disclosed in the present application is a newly identifiedLDL receptor homolog.

[1166] 29. Full-Length PRO772 Polypeptides

[1167] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO772. In particular, Applicants have identified andisolated cDNA encoding a PRO772 polypeptide, as disclosed in fer detailin the Examples below. Using BLAST and FastA sequence alignment computerprograms, Applicants found that the PRO772 polypeptide has significantsimilarity to the human A4 protein. Accordingly, it is presentlybelieved that PRO772 polypeptide disclosed in the present application isa newly identified A4 protein homolog.

[1168] 30. Full-Length PRO852 Polypeptides

[1169] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO852. In particular, Applicants have identified andisolated cDNA encoding a PRO852 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO852 polypeptide hassignificant similarity to various protease proteins. Accordingly, it ispresently believed that PRO852 polypeptide disclosed in the presentapplication is a newly identified protease enzyme homolog.

[1170] 31. Full-Length PRO853 Polypeptides

[1171] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO853. In particular, Applicants have identified andisolated cDNA encoding a PRO853 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO853 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO853polypeptide possess significant sequence similarity to the reductaseprotein, thereby indicating that PRO853 may be a novel reductase. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant sequence similarity between thePRO853 amino acid sequence and the following Dayhoff sequences,“P_W03198”, “CEC15H11_(—)6”, “MTV030_(—)12”, “P_W15759”, “S42651”,“ATAC00234314”, “MTV022_(—)13”, “SCU437041”, “CELE04F6_(—)7”, and“ALFA_(—)1”. Accordingly, it is presently believed that PRO853polypeptide disclosed in the present application is a newly identifiedmember of the reductase family and possesses the antioxidant enzymaticactivity typical of the reductase family.

[1172] 32. Full-Length PRO860 Polypeptides

[1173] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO860. In particular, Applicants have identified andisolated cDNA encoding a PRO860 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO860 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO860polypeptide possess significant sequence similarity to the neurofascinprotein, thereby indicating that PRO860 may be a novel neurofascin. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant sequence similarity between thePRO860 amino acid sequence and the following Dayhoff sequences,“AF040990_(—)1”, “AF041053_(—)1”, “CELZK377_(—)2”, “RNU81035_(—)1”,“D86983_(—)1”, “S26180”, “MMBIG2A_(—)1”, “S46216”, and “RNU68726_(—)1”.Accordingly, it is presently believed that PRO860 polypeptide disclosedin the present application is a newly identified member of theneurofascin family and possesses the cellular adhesion propertiestypical of the neurofascin family.

[1174] 33. Full-Length PRO846 Polypeptides

[1175] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO846. In particular, Applicants have identified andisolated cDNA encoding a PRO846 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO846 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO846polypeptide possess significant sequence similarity to the CMRF35protein, thereby indicating that PRO846 may be a novel CMRF35 protein.More specifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant sequence similarity between thePRO846 amino acid sequence and the following Dayhoff sequences,“CM35_HUMAN”, “AF035963_(—)1”, “PIGR_RABIT”, “AF043724_(—)1”,“RNU89744_(—)1”, “A52091_(—)1”, “S48841”, “ELK06A9_(—)3”, and“AF049588_(—)1”. Accordingly, it is presently believed that PRO846polypeptide disclosed in the present application is a newly identifiedmember of the CMRF35 protein family and possesses properties typical ofthe CMRF35 protein family.

[1176] 34. Full-Length PRO862 Polypeptides

[1177] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO862. In particular, Applicants have identified andisolated cDNA encoding a PRO862 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO862 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO862polypeptide possess significant sequence similarity to the lysozymeprotein, thereby indicating that PRO862 may be a novel lysozyme protein.More specifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant sequence similarity between thePRO862 amino acid sequence and the following Dayhoff sequences,“P_P90343”, and “LYC_HUMAN”. Accordingly, it is presently believed thatPRO862 polypeptide disclosed in the present application is a newlyidentified member of the lysozyme family and possesses catalyticactivity typical of the lysozyme family.

[1178] 35. Full-Length PRO864 Polypeptides

[1179] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO864. In particular, Applicants have identified andisolated cDNA encoding a PRO864 polypeptide, as disclosed in furtherdetail in the Examples below. Analysis of the amino acid sequence of thefull-length PRO864 polypeptide using BLAST and FastA sequence alignmentcomputer programs, suggests that various portions of the PRO864polypeptide possess significant sequence similarity to the Wnt-4protein, thereby indicating that PRO864 may be a novel Wnt-4 protein.More specifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant sequence similarity between thePRO864 amino acid sequence and the following Dayhoff sequences,“WNT4_MOUSE”, “WNT3_MOUSE”, “WN5A_HUMAN”, “WN7B_MOUSE”, “WN3A_MOUSE”,“XLU66288_(—)1”, “WN13_HUMAN”, “WN5B_ORYLA”, “WNT2_MOUSE”, and“WN7A_MOUSE”. Accordingly, it is presently believed that PRO864polypeptide disclosed in the present application is a newly identifiedmember of the Wnt-4 protein family and possesses properties typical ofthe Wnt-4 protein family.

[1180] 36. Full-Length PRO792 Polypeptides

[1181] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO792. hI particular, Applicants have identified andisolated cDNA a, encoding a PRO792 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO792 polypeptide hassignificant similarity to the CD23 protein. Accordingly, it is presentlybelieved that PRO792 polypeptide disclosed in the present application isa newly identified CD23 homolog.

[1182] 37. Full-Length PRO866 Polypeptides

[1183] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO866. In particular, Applicants have identified andisolated cDNA encoding a PRO866 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO866 polypeptide hassignificant similarity to various mindin and spondin proteins.Accordingly, it is presently believed that PRO866 polypeptide disclosedin the present application is a newly identified mindin/spondin homolog.

[1184] 38. Full-Length PRO871 Polypeptides

[1185] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO871. Ia particular, Applicants have identified andisolated cDNA encoding a PRO871 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO871 polypeptide hassignificant similarity to the CyP-60 protein. Accordingly, it ispresently believed that PRO871 polypeptide disclosed in the presentapplication is a newly identified member of the cyclophilin proteinfamily and possesses activity typical of the cyclophilin protein family.

[1186] 39. Full-Length PRO873 Polypeptides

[1187] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO873. In particular, Applicants have identified andisolated cDNA encoding a PRO873 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO873 polypeptide hassignificant similarity to a human liver carboxylesterase. Accordingly,it is presently believed that PRO873 polypeptide disclosed in thepresent application is a newly identified member of the carboxylesterasefamily and possesses enzymatic activity typical of the carboxylesterasefamily.

[1188] 40. Full-Length PRO940 Polypeptides

[1189] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO940. In particular, Applicants have identified andisolated cDNA encoding a PRO940 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO940 polypeptide hassignificant similarity to CD33 and the OB binding protein-2.Accordingly, it is presently believed that PRO940 polypeptide disclosedin the present application is a newly CD33 and/or OB binding protein-2homolog.

[1190] 41. Full-Length PRO941 Polypeptides

[1191] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO941. In particular, Applicants have identified andisolated cDNA encoding a PRO941 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO941 polypeptide hassignificant similarity to one or more cadherin proteins. Accordingly, itis presently believed that PRO941 polypeptide disclosed in the presentapplication is a newly identified cadherin homolog.

[1192] 42. Full-Length PRO944 Polypeptides

[1193] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO944. In particular, Applicants have identified andisolated cDNA encoding a PRO944 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO944 polypeptide hassignificant similarity to the CPE-R cell surface protein. Accordingly,it is presently believed that PRO944 polypeptide disclosed in thepresent application is a newly identified CPE-R homolog.

[1194] 43. Full-Length PRO983 Polypeptides

[1195] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO983. In particular, Applicants have identified andisolated cDNA encoding a PRO983 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO983 polypeptide hassignificant similarity to the vesicle-associated protein, VAP-33.Accordingly, it is presently believed that PRO983 polypeptide disclosedin the present application is a newly identified member of thevesicle-associated membrane protein family and possesses activitytypical of vesicle-associated membrane proteins.

[1196] 44. Full-Length PRO1057 Polypeptides

[1197] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1057. In particular, Applicants have identified andisolated cDNA encoding a PRO1057 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO1057 polypeptide hassignificant similarity to various protease proteins. Accordingly, it ispresently believed that PRO1057 polypeptide disclosed in the presentapplication is a newly identified protease homolog.

[1198] 45. Full-Length PRO1071 Polypeptides

[1199] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1071. In particular, Applicants have identified andisolated cDNA encoding a PRO1071 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO1071 polypeptide hassignificant similarity to the thrombospondin protein. Accordingly, it ispresently believed that PRO1071 polypeptide disclosed in the presentapplication is a newly identified thrombospondin homolog.

[1200] 46. Full-Length PRO1072 Polypeptides

[1201] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1072. In particular, Applicants have identified andisolated cDNA encoding a PRO1072 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO1072 polypeptide hassignificant similarity to various reductase proteins. Accordingly, it ispresently believed that PRO1072 polypeptide disclosed in the presentapplication is a newly identified member of the reductase proteinfamily.

[1202] 47. Full-Length PRO1075 Polypeptides

[1203] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1075. In particular, Applicants have identified andisolated cDNA encoding a PRO1075 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO1075 polypeptide hassignificant similarity to protein disulfide isomerase. Accordingly, itis presently believed that PRO1075 polypeptide disclosed in the presentapplication is a newly identified member of the protein disulfideisomerase family and possesses activity typical of that family.

[1204] 48. Full-Length PRO181 Polypeptides

[1205] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO181. In particular, Applicants have identified andisolated cDNA encoding a PRO181 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO181 polypeptide hassignificant similarity to the cornichon protein. Accordingly, it ispresently believed that PRO181 polypeptide disclosed in the presentapplication is a newly identified cornichon homolog.

[1206] 49. Full-Length PRO195 Polypeptides

[1207] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO195. In particular, Applicants have identified andisolated cDNA encoding a PRO195 polypeptide, as disclosed in furtherdetail in the Examples below. The PRO195-encoding clone was isolatedfrom a human fetal placenta library using a trapping technique whichselects for nucleotide sequences encoding secreted proteins. ToApplicants present knowledge, the UNQ169 (DNA26847-1395) nucleotidesequence encodes a novel factor; using BLAST and FastA sequencealignment computer programs, no sequence identities to any knownproteins were revealed.

[1208] 50. Full-Length PRO865 Polypeptides

[1209] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO865. In particular, Applicants have identified andisolated cDNA encoding a PRO865 polypeptide, as disclosed in furtherdetail in the Examples below. The PRO865-encoding clone was isolatedfrom a human fetal kidney library using a trapping technique whichselects for nucleotide sequences encoding secreted proteins. Thus, thePRO865-encoding clone may encode a secreted factor. To Applicantspresent knowledge, the UNQ434 (DNA53974-1401) nucleotide sequenceencodes a novel factor; using BLAST and FastA sequence alignmentcomputer programs, no sequence identities to any known proteins wererevealed.

[1210] 51. Full-Length PRO827 Polypeptides

[1211] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO827. In particular, Applicants have identified andisolated cDNA encoding a PRO827 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO827 polypeptide hassignificant similarity to VLA-2 and various other integrin proteins.Accordingly, it is presently believed that PRO827 polypeptide disclosedin the present application is a novel integrin protein or splice variantthereof.

[1212] 52. Full-Length PRO1114 Polypeptides

[1213] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1114. In particular, Applicants have identified andisolated cDNA encoding a PRO1114 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO1114 polypeptide hassignificant similarity to the cytokine receptor family of proteins.Accordingly, it is presently believed that PRO1114 polypeptide disclosedin the present application is a newly identified member of the cytokinereceptor family of proteins and possesses activity typical of thatfamily.

[1214] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1114 interferon receptor (UNQ557). In particular, cDNAencoding a PRO1114 interferon receptor polypeptide has been identifiedand isolated, as disclosed in further detail in the Examples below. Itis noted that proteins produced in separate expression rounds may begiven different PRO numbers but the UNQ number is unique for any givenDNA and the encoded protein, and will not be changed. However, for sakeof simplicity, in the present specification the protein encoded byDNA57033-1403 as well as all further native homologues and variantsincluded in the foregoing definition of PRO1114 interferon receptor,will be referred to as “PRO1114 interferon receptor”, regardless oftheir origin or mode of preparation.

[1215] Using the WU-BLAST2 sequence alignment computer program, it hasbeen found that a full-length native sequence PRO1114 interferonreceptor polypeptide (shown in FIG. 142 and SEQ ID NO:352) has sequenceidentity with the other known interferon receptors. Accordingly, it ispresently believed that PRO1114 interferon receptor possesses activitytypical of other interferon receptors.

[1216] 53. Full-Length PRO237 Polypeptides

[1217] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO237. In particular, Applicants have identified andisolated cDNA encoding a PRO237 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO237 polypeptide hassignificant similarity to carbonic anhydrase. Accordingly, it ispresently believed that PRO237 polypeptide disclosed in the presentapplication is a newly identified carbonic anhydrase homolog.

[1218] 54. Full-Length PRO541 Polypeptides

[1219] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO541. In particular, Applicants have identified andisolated cDNA encoding a PRO541 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO541 polypeptide hassignificant similarity to a trypsin inhibitor protein. Accordingly, itis presently believed that PRO541 polypeptide disclosed in the presentapplication is a newly identified member of the trypsin inhibitorprotein family.

[1220] 55. Full-Length PRO273 Polypeptides

[1221] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO273. In particular, Applicants have identified andisolated cDNA encoding a PRO273 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO273polypeptide have significant sequence identity with various chemokines.Accordingly, it is presently believed that PRO273 polypeptide disclosedin the present application is a newly identified member of the chemokinefamily and possesses activity typical of the chemokine family.

[1222] 56. Full-Length PRO701 Polypeptides

[1223] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO701. In particular, Applicants have identified andisolated cDNA encoding a PRO701 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO701polypeptide have significant homology with the neuroligins 1, 2 and 3and esterases including carboxyesterases and acytlcholinesterases.Accordingly, it is presently believed that PRO701 polypeptide disclosedin the present application is a newly identified member of theneuroligin family and is involved in mediating recognition processesbetween neurons and/or functions as a cell adhesin molecule as istypical of neuroligins.

[1224] 57. Full-Length PRO704 Polypeptides

[1225] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO704. In particular, Applicants have identified andisolated cDNA encoding a PRO704 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO704polypeptide have significant homology with the VIP36 and GP36b.Accordingly, it is presently believed that PRO704 polypeptide disclosedin the present application is a newly identified member of the vesicularintegral membrane protein family and possesses the ability to bind tosugars and cycle between the plasma membrane and the Golgi typical ofthis family.

[1226] 58. Full-Length PRO706 Polypeptides

[1227] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO706. In particular, Applicants have identified andisolated cDNA encoding a PRO706 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO706polypeptide have sequence identity with the human prostatic acidphosphatase precursor and the human lysosomal acid phosphataseprecursor. Accordingly, it is presently believed that PRO706 polypeptidedisclosed in the present application is a newly identified member of thehuman prostatic acid phosphatase precursor family and possessesphosphatase typical of the acid phosphatase family.

[1228] 59. Full-Length PRO707 Polypeptides

[1229] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO707. In particular, Applicants have identified andisolated cDNA encoding a PRO707 polypeptide, as disclosed in futherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO707polypeptide have significant homology with cadherins, particularlycadherin FIB3 found in fibroblasts. Accordingly, it is presentlybelieved that PRO707 polypeptide disclosed in the present application isa newly identified member of the cadherin family and possesses cellinteraction signaling typical of the cadherin family.

[1230] 60. Full-Length PRO322 Polypeptides

[1231] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO322. In particular, Applicants have identified andisolated cDNA encoding a PRO322 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO322polypeptide have significant homology with human neuropsin, serineprotease, neurosin and trypsinogen. Accordingly, it is presentlybelieved that PRO322 polypeptide disclosed in the present application isa newly identified member of the serine protease family and possessesprotease activity typical of this family. It is also believed thatPRO322 is involved in hippocampal plasticity and is associated withextracellular matrix modifications and cell migrations.

[1232] 61. Full-Length PRO526 Polypeptides

[1233] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO526. In particular, Applicants have identified andisolated cDNA encoding a PRO526 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO526polypeptide have significant homology with the acid labile subunit ofthe insulin-like growth factor complex (ALS), as well carboxypeptidase,SLIT, and platelet glycoprotein V. Accordingly, it is presently believedthat PRO526 polypeptide disclosed in the present application is a newlyidentified member of the leucine-repeat rich superfamily, and possessesprotein-protein interaction capabilities typical of this family.

[1234] 62. Full-Length PRO531 Polypeptides

[1235] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO531. In particular, Applicants have identified andisolated cDNA encoding a PRO531 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO531polypeptide have significant sequence identity and similarity withmembers of the cadherin superfamily, particularly, protocadherinAccordingly, it is presently believed that PRO531 polypeptide disclosedin the present application is a newly identified member of the cadherinsuperfamily, and is a protocadherin. PRO531 is a transmembrane proteinwhich has extracellular cadherin motifs. PRO531 is believed to beinvolved in cell-cell activity, in particular, cell signaling.

[1236] 63. Full-Length PRO534 Polypeptides

[1237] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO534. In particular, Applicants have identified andisolated cDNA encoding a PRO534 polypeptide, as disclosed in futherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO534polypeptide have significant identity or similarity with the putativedisulfide isomerase erp38 precursor and thioredoxin c-3. Accordingly, itis presently believed that PRO534 polypeptide disclosed in the presentapplication is a newly identified member of the disulfide isomerasefamily and possesses the ability to recognize and unscramble eitherintermediate or incorrect folding patterns typical of this family.

[1238] 64. Full-Length PRO697 Polypeptides

[1239] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO697. In particular, Applicants have identified andisolated cDNA encoding a PRO697 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO697polypeptide have significant identity or similarity with sFRP-2, sFRP-1and SARP-1, -2 and -3. Accordingly, it is presently believed that PRO697polypeptide disclosed in the present application is a newly identifiedmember of the sFRP family and possesses activity related to the Wntsignal pathway.

[1240] 65. Full-Length PRO717 Polypeptides

[1241] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO717. In particular, Applicants have identified andisolated cDNA encoding a PRO717 polypeptide, as disclosed in furtherdetail in the Examples below. To Applicants present knowledge, theUNQ385 (DNA50988-1326) nucleotide sequence encodes a novel factor; usingBLAST and FastA sequence alignment computer programs, no significantsequence identities to any known human proteins were revealed.

[1242] 66. Full-Length PRO731 Polypeptides

[1243] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO731. In particular, Applicants have identified andisolated cDNA encoding a PRO731 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various positions of the PRO731polypeptide have significant homology with the protocadherins 4, 68, 43,42, 3, and 5. Accordingly, it is presently believed that PRO731polypeptide disclosed in the present application is a newly identifiedmember of the protocadherin family and possesses cell-cell aggregationor signaling activity or signal transduction involvement typical of thisfamily.

[1244] 67. Full-length PRO218 Polypeptides

[1245] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO218. In particular, Applicants have identified andisolated cDNA encoding a PRO218 polypeptide, as disclosed in furtherdetail in the Examples below. The PRO218-encoding clone was isolatedfrom a human fetal kidney library. To Applicants present knowledge, theUNQ192 (DNA30867-1335) nucleotide sequence encodes a novel factor; usingBLAST and FastA sequence alignment computer programs, no significantsequence identities to any known proteins were revealed. Some sequenceidentity was found with membrane regulator proteins, indicating thatPRO218 may function as a membrane regulator.

[1246] 68. Full-Length PRO768 Polypeptides

[1247] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO768. In particular, Applicants have identified andisolated cDNA encoding a PRO768 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO768polypeptide have significant homology with integrins, including integrin7 and 6. Accordingly, it is presently believed that PRO768 polypeptidedisclosed in the present application is a newly identified member of theintegrin family, either a homologue or a splice variant of integrin 7,and is involved with cell adhesion and communication between musclecells and the extracellular matrix.

[1248] 69. Full-Length PRO771 Polypeptides

[1249] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO771. In particular, Applicants have identified andisolated cDNA encoding a PRO771 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO771polypeptide have significant sequence identity and similarity withtestican. Accordingly, it is presently believed that PRO771 polypeptidedisclosed in the present application is a newly identified member of thetestican family and possesses cell signaling, binding, or adhesionproperties, typical of this family.

[1250] 70. Full-Length PRO733 Polypeptides

[1251] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO733. In particular, Applicants have identified andisolated cDNA encoding a PRO733 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO733polypeptide nave significant sequence identity with the T1/ST receptorbinding protein. Accordingly, it is presently believed that PRO733polypeptide disclosed in the present application is a newly identifiedmember of the interleulin-like family binding proteins which may be acytokine and which may be involved in cell signaling. It is believedthat PRO733 is an ApoAIV homologue.

[1252] 71. Full-Length PRO162 Polypeptides

[1253] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO162. In particular, Applicants have identified andisolated cDNA encoding a PRO162 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO162polypeptide have significant homology with human pancreatitis-associatedprotein (PAP). Applicants have also found that the DNA encoding thePRO162 polypeptide has significant homology with bovine lithostathineprecursor and bovine pancreatic thread protein (PTP). Accordingly, it ispresently believed that PRO162 polypeptide disclosed in the presentapplication is a newly identified member of the pancreatitis-associatedprotein family and possesses activity typical of thepancreatitis-associated protein family.

[1254] 72. Full-Length PRO788 Polypeptides

[1255] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO788. In particular, Applicants have identified andisolated cDNA encoding a PRO788 polypeptide, as disclosed in fintherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO788polypeptide have significant homology with the anti-neoplastic urinaryprotein. Applicants have also found that the DNA encoding the PRO788polypeptide has significant homology with human E48 antigen, humancomponent B protein, and human prostate stem cell antigen. Accordingly,it is presently believed that PRO788 polypeptide disclosed in thepresent application is a newly identified member of the anti-neoplasticurinary protein family and possesses anti-neoplastic activity typical ofthe anti-neoplastic urinary protein family.

[1256] 73. Full-Length PRO1008 Polypeptides

[1257] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1008. in particular, Applicants have identified andisolated cDNA encoding a PRO1008 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO1008polypeptide have significant sequence identity and similarity with mousedkk-1 (mdkk-1). Accordingly, it is presently believed that PRO1008polypeptide disclosed in the present application is a newly identifiedmember of the dkk-1 family and possesses head inducing activity typicalof this family.

[1258] 74. Full-Length PRO1012 Polypeptides

[1259] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1012. In particular, Applicants have identified andisolated cDNA encoding a PRO1012 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO1012polypeptide have sequence identity with disulfide isomerase.Accordingly, it is presently believed that PRO1012 polypeptide disclosedin the present application is a newly identified member of the ERretained protein family and possesses activity related to theprocessing, production and/or folding of polypeptides typical of thedisulfide isomerase family.

[1260] 75. Full-Length PRO1014 Polypeptides

[1261] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1014. In particular, Applicants have identified andisolated cDNA encoding a PRO1014 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO1014polypeptide have sequence identity with reductase and dehydrogenase.Accordingly, it is presently believed that PRO1014 polypeptide disclosedin the present application is a newly identified member of the reductasesuper family and possesses reduction capabilities typical of thisfamily.

[1262] 76. Full-Length PRO1017 Polypeptides

[1263] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1017. In particular, Applicants have identified andisolated cDNA encoding a PRO1017 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO1017polypeptide have sequence identity with HNK-1 sulfotransferase.Accordingly, it is presently believed that PRO1017 polypeptide disclosedin the present application is a newly identified member of the HNK-1sulfotransferase family and is involved with the synthesis of HNK-1carbohydrate epitopes typical of this family.

[1264] 77. Full-Length PRO474 Polypeptides

[1265] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO474. In particular, Applicants have identified andisolated cDNA encoding a PRO474 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO474polypeptide have sequence identity with dehydrogenase, glucosedehydrogenase and oxidoreductase. Accordingly, it is presently believedthat PRO474 polypeptide disclosed in the present application is a newlyidentified member of the dehydrogenase family and is involved in theoxidation of glucose.

[1266] 78. Full-length PRO1031 Polypeptides

[1267] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1031. In particular, Applicants have identified andisolated cDNA encoding a PRO1031 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO1031polypeptide have sequence identity with IL-17 and CTLA-8. Accordingly,it is presently believed that PRO1031 polypeptide disclosed in thepresent application is a newly identified member of the cytokine familyand thus may be involved in inflammation and/or the immune system.

[1268] 79. Full-Length PRO938 Polypeptides

[1269] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO938. In particular, Applicants have identified andisolated cDNA encoding a PRO938 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO938 polypeptide hassignificant similarity to protein disulfide isomerase. Accordingly, itis presently believed that PRO938 polypeptide disclosed in the presentapplication is a newly identified member of the thioredoxin familyproteins and possesses activity typical of protein disulfide isomerase.

[1270] 80. Full-Length PRO1082 Polypeptides

[1271] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1082. In particular, Applicants have identified andisolated cDNA encoding a PRO1082 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that various portions of the PRO1082polypeptide have sequence identity with a lectin-like oxidized LDLreceptor appearing in the database as “AB010710_(—)1”. Accordingly, itis presently believed that PRO1082 polypeptide disclosed in the presentapplication is a newly identified member of the LDL receptor family.

[1272] 81. Full-Length PRO1083 Polypeptides

[1273] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1083. In particular, Applicants have identified andisolated cDNA encoding a PRO1083 polypeptide, as disclosed in furtherdetail in the Examples below. The PRO1083-encoding clone was isolatedfrom a human fetal kidney library using a trapping technique whichselects for nucleotide sequences encoding secreted proteins. ToApplicants present knowledge, the UNQ540 (DNA50921-1458) nucleotidesequence encodes a novel factor; using BLAST and FastA sequencealignment computer programs, some sequence identity with a 7TM receptor,latrophilin related protein 1 and a macrophage restricted cell surfaceglycoprotein was shown. The kinase phosphorylation site and G-coupledreceptor domain shown in FIG. 204 indicate that PRO1083 is a novelmember of the 7TM receptor superfamily.

[1274] 82. Full-Length PRO200 Polypeptides

[1275] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as VEGF-E. In particular, Applicants have identified andisolated cDNA encoding a VEGF-E polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST sequence alignment computerprograms, Applicants found that the VEGF-E polypeptide has significanthomology with VEGF and bone morphogenetic protein 1. In particular, thecDNA sequence of VEGF-E exhibits 24% amino acid similarity with VEGF andhas structural conservation. In addition, VEGF-E contains a N-terminalhalf which is not present in VEGF and that has 28% homology to bonemorphogenetic protein 1.

[1276] 83. Full-Length PRO285 and PRO286 Polypeptides

[1277] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO285 and PRO286 In particular, Applicants haveidentified and isolated cDNAs encoding PRO285 and PRO286 polypeptides,as disclosed in further detail in the Examples below. Using BLAST andFastA sequence alignment computer programs, Applicants found that thecoding sequences of PRO285 and PRO286 are highly homologous to DNAsequences HSU88540_(—)1, HSU88878_(—)1, HSU88879_(—)1, HSU88880_(—)1,and HSU88881_(—)1 in the GenBank database.

[1278] Accordingly, it is presently believed that the PRO285 and PRO286proteins disclosed in the present application are newly identified humanhomologues of the Drosophila protein Toll, and are likely to play animportant role in adaptive immunity. More specifically, PRO285 andPRO286 may be involved in inflammation, septic shock, and response topathogens, and play possible roles in diverse medical conditions thatare aggravated by immune response, such as, for example, diabetes, ALS,cancer, rheumatoid arthritis, and ulcers. The role of PRO285 and PRO286as pathogen pattern recognition receptors, sensing the presence ofconserved molecular structures present on microbes, is further supportedby the data disclosed in the present application, showing that a knownhuman Toll-like receptor, TLR2 is a direct mediator of LPS signaling.

[1279] 84. Full-Length PRO213-1. PRO1330 and PRO1449 Polypeptides

[1280] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO213-1, PRO1330 and/or PRO1449. In particular, cDNAencoding a PRO213-1, PRO1330 and/or PRO1449 polypeptide has beenidentified and isolated, as disclosed in further detail in the Examplesbelow. It is noted that proteins produced in separate expression roundsmay be given different PRO numbers but the UNQ number is unique for anygiven DNA and the encoded protein, and will not be changed. However, forsake of simplicity, in the present specification the protein encoded byDNA30943-1163-1, DNA64907-1163-1 and DNA64908-1163-1 as well as allfurther native homologues and variants included in the foregoingdefinition ofPRO213-1, PRO1330 and/or PRO1449, will be referred to as“PRO213-1, PRO1330 and/or PRO1449”, regardless of their origin or modeof preparation.

[1281] 85. Full-Length PRO298 Polypeptides

[1282] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO298. (It is noted that PRO298 is an arbitrarydesignation of a protein encoded by the nucleic acid shown in FIG. 218,SEQ ID NO:514, and having the amino acid sequence shown in FIG. 219, SEQID NO:515. Further proteins having the same amino acid sequence butexpressed in different rounds of expression, may be given different“PRO” numbers.) In particular, Applicants have identified and isolatedcDNA encoding a PRO298 polypeptide, as disclosed in further detail inthe Examples below. Using BLASIX 2.0a8MP-WashU computer program, socringparameters: T=12, S=68, S2=36, Matrix: BLOSUM62, Applicants found thatthe PRO298 protein specifically disclosed herein shows a limited(27-38%) sequence identity with the following sequences found in theGenBank database: S59392 (probable membrane protein YLR246w—yeast);S58154 hypothetical protein SPAC2F7.10—yeast); CELF33D11_(—)9(F33D11.9b—Caenorhabditis elegans); YO41_CAEEL (hypothetical 68.7 kdprotein zk757.1); CEAC3_(—)5 (AC3.4—Caenorhabditis elegans); S52691(probable transmembrane protein YDR126w—yeast); ATT12H17_(—)14(protein—Arabidopsis thaliana); S55963 (probable membrane proteinYNL326c—yeast); CELC43H6_(—)2 (C43H6.7—Caenorhabditis elegans);TMO18A10_(—)14 (A_TMO18A10.8—Arabinosa thaliana).

[1283] 86. Full-Length PRO337 Polypeptides

[1284] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO337. In particular, Applicants have identified andisolated cDNA encoding a PRO337 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST, BLAST-2 and FastA sequencealignment computer programs, Applicants found that a full-length nativesequence PRO337 has 97% amino acid sequence identity with rat neurotrin,85% sequence identity with chicken CEPU, 73% sequence identity withchicken G55, 59% homology with human LAMP and 84% homology with humanOPCAM. Accordingly, it is presently believed that PRO337 disclosed inthe present application is a newly identified member of the IgLON subfamily of the immunoglobulin superfamily and may possess neurite growthand differentiation potentiating properties.

[1285] 87. Full-Length PRO403 Polypeptides

[1286] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO403. In particular, Applicants have identified andisolated cDNA encoding a PRO403 polypeptide, as disclosed in furtherdetail in the Examples below. Using a BLAST, BLAST-2 and FastA sequencealignment computer programs, Applicants found that a full-length nativesequence PRO403 has 94% identity to bovine ECE-2 and 64% identity tohuman ECE-1. Accordingly is presently believed that PRO403 is a newmember of the ECE protein family and may posses ability to catalyze theproduction of active endothelin.

[1287] B. PRO Polypeptide Variants

[1288] In addition to the full-length native sequence PRO polypeptidesdescribed herein, it is contemplated that PRO variants can be prepared.PRO variants can be prepared by introducing appropriate nucleotidechanges into the PRO DNA, and/or by synthesis of the desired PROpolypeptide. Those skilled in the art will appreciate that amino acidchanges may alter post-translational processes of the PRO, such aschanging the number or position of glycosylation sites or altering themembrane anchoring characteristics.

[1289] Variations in the native full-length sequence PRO or in variousdomains of the PRO described herein, can be made, for example, using anyof the techniques and guidelines for conservative and non-conservativemutations set forth, for instance, in U.S. Pat. No. 5,364,934.Variations may be a substitution, deletion or insertion of one or morecodons encoding the PRO that results in a change in the amino acidsequence of the PRO as compared with the native sequence PRO. Optionallythe variation is by substitution of at least one amino acid with anyother amino acid in one or more of the domains of the PRO. Guidance indetermining which amino acid residue may be inserted, substituted ordeleted without adversely affecting the desired activity may be found bycomparing the sequence of the PRO with that of homologous known proteinmolecules and minimizing the number of amino acid sequence changes madein regions of high homology. Amino acid substitutions can be the resultof replacing one amino acid with another amino acid having similarstructural and/or chemical properties, such as the replacement of aleucine with a serine, i.e., conservative amino acid replacements.Insertions or deletions may optionally be in the range of about 1 to 5amino acids. The variation allowed may be determined by systematicallymaking insertions, deletions or substitutions of amino acids in thesequence and testing the resulting variants for activity exhibited bythe full-length or mature native sequence.

[1290] PRO polypeptide fragments are provided herein. Such fragments maybe truncated at the N-terminus or C-terminus, or may lack internalresidues, for example, when compared with a full length native protein.Certain fragments lack amino acid residues that are not essential for adesired biological activity of the PRO polypeptide.

[1291] PRO fragments may be prepared by any of a number of conventionaltechniques. Desired peptide fragments may be chemically synthesized. Analternative approach involves generating PRO fragments by enzymaticdigestion, e.g., by treating the protein with an enzyme known to cleaveproteins at sites defined by particular amino acid residues, or bydigesting the DNA with suitable restriction enzymes and isolating thedesired fragment. Yet another suitable technique involves isolating andamplifying a DNA fragment encoding a desired polypeptide fragment, bypolymerase chain reaction (PCR). Oligonucleotides that define thedesired termini of the DNA fragment are employed at the 5′ and 3′primers in the PCR. Preferably, PRO polypeptide fragments share at leastone biological and/or immunological activity with the native PROpolypeptide disclosed herein.

[1292] In particular embodiments, conservative substitutions of interestare shown in Table 6 under the heading of preferred substitutions. Ifsuch substitutions result in a change in biological activity, then moresubstantial changes, denominated exemplary substitutions in Table 6, oras further described below in reference to amino acid classes, areintroduced and the products screened. TABLE 6 Original ExemplaryPreferred Residue Substitutions Substitutions Ala (A) val; leu; ile valArg (R) lys; gln; asn lys Asn (N) gln; his; lys; arg gln Asp (D) glu gluCys (C) ser ser Gln (Q) asn asn Glu (E) asp asp Gly (G) pro; ala ala His(H) asn; gln; lys; arg arg Ile (I) leu; val; met; ala; phe; leunorleucine Leu (L) norleucine; ile; val; ile met; ala; phe Lys (K) arg;gln; asn arg Met (M) leu; phe; ile leu Phe (F) leu; val; ile; ala; tyrleu Pro (P) ala ala Ser (S) thr thr Thr (T) ser ser Trp (W) tyr; phe tyrTyr (Y) trp; phe; thr; ser phe Val (V) ile; leu; met; phe; leu ala;norleucine

[1293] Substantial modifications in function or immunological identityof the PRO polypeptide are accomplished by selecting substitutions thatdiffer significantly in their effect on maintaining (a) the structure ofthe polypeptide backbone in the area of the substitution, for example,as a sheet or helical conformation, (b) the charge or hydrophobicity ofthe molecule at the target site, or (c) the bulk of the side chain.Naturally occurring residues are divided into groups based on commonside-chain properties:

[1294] (1) hydrophobic: norleucine, met, ala, val, leu, ile;

[1295] (2) neutral hydrophilic: cys, ser, thr;

[1296] (3) acidic: asp, gIu;

[1297] (4) basic: asn, gln, his, lys, arg;

[1298] (5) residues that influence chain orientation: gly, pro; and

[1299] (6) aromatic: trp, tyr, phe.

[1300] Non-conservative substitutions will entail exchanging a member ofone of these classes for another class. Such substituted residues alsomay be introduced into the conservative substitution sites or, morepreferably, into the remaining (non-conserved) sites.

[1301] The variations can be made using methods known in the art such asoligonucleotide-mediated (site-directed) mutagenesis, alanine scanning,and PCR mutagenesis. Site-directed mutagenesis [Carter et al., Nucl.Acids Res., 13:4331 (1986); Zoller et al., Nucl. Acids Res. 10:6487(1987)], cassette mutagenesis [Wells et al., Gene, 34:315 (1985)],restriction selection mutagenesis [Wells et al., Philos. Trans. R. Soc.London SerA, 317:415 (1986)] or other known techniques can be performedon the cloned DNA to produce the PRO variant DNA.

[1302] Scanning amino acid analysis can also be employed to identify oneor more amino acids along a contiguous sequence. Among the preferredscanning amino acids are relatively small, neutral amino acids. Suchamino acids include alanine, glycine, serine, and cysteine. Alanine istypically a preferred scanning amino acid among this group because iteliminates the side-chain beyond the beta-carbon and is less likely toalter the main-chain conformation of the variant [Cunningham and Wells,Science, 244: 1081-1085 (1989)]. Alanine is also typically preferredbecause it is the most common amino acid. Further, it is frequentlyfound in both buried and exposed positions [Creighton, The Proteins,(W.H. Freeman & Co., N.Y.); Chothia, J. Mol. Biol., 150:1 (1976)]. Ifalanine substitution does not yield adequate amounts of variant, anisoteric amino acid can be used.

[1303] C. Modifications of PRO

[1304] Covalent modifications of PRO are included within the scope ofthis invention. One type of covalent modification includes reactingtargeted amino acid residues of a PRO polypeptide with an organicderivatizing agent that is capable of reacting with selected side chainsor the N- or C- terminal residues of the PRO. Derivatization withbifunctional agents is useful, for instance, for crosslinking PRO to awater-insoluble support matrix or surface for use in the method forpurifying anti-PRO antibodies, and vice-versa. Commonly usedcrosslinking agents include, e.g., 1,1-bis(diazoacetyl)-2-phenylethane,glutaraldehyde, N-hydroxysuccinimide esters, for example, esters with4-azidosalicylic acid, homobifunctional imidoesters, includingdisuccininidyl esters such as 3,3′-dithiobis(succinimidylpropionate),bifunctional maleimides such as bis-N-maleimido-1,8-octane and agentssuch as methyl-3-[(pazidophenyl)dithiolpropioimidate.

[1305] Other modifications include deamidation of glutaminyl andasparaginyl residues to the corresponding glutamyl and aspartylresidues, respectively, hydroxylation of proline and lysine,phosphorylation of hydroxyl groups of seryl or threonyl residues,methylation of the α-amino groups of lysine, arginine, and histidineside chains [T. E. Creighton, Proteins: Structure and MolecularProperties, W.H. Freeman & Co., San Francisco, pp. 79-86 (1983)],acetylation of the N-terminal amine, and amidation of any C-terminalcarboxyl group.

[1306] Another type of covalent modification of the PRO polypeptideincluded within the scope of this invention comprises altering thenative glycosylation pattern of the polypeptide. “Altering the nativeglycosylation pattern” is intended for purposes herein to mean deletingone or more carbohydrate moieties found in native sequence PRO (eitherby removing the underlying glycosylation site or by deleting theglycosylation by chemical and/or enzymatic means), and/or adding one ormore glycosylation sites that are not present in the native sequencePRO. In addition, the phrase includes qualitative changes in theglycosylation of the native proteins, involving a change in the natureand proportions of the various carbohydrate moieties present.

[1307] Addition of glycosylation sites to the PRO polypeptide may beaccomplished by altering the amino acid sequence. The alteration may bemade, for example, by the addition of, or substitution by, one or moreserine or threonine residues to the native sequence PRO (for O-linkedglycosylation sites). The PRO amino acid sequence may optionally bealtered through changes at the DNA level, particularly by mutating theDNA encoding the PRO polypeptide at preselected bases such that codonsare generated that will translate into the desired amino acids.

[1308] Another means of increasing the number of carbohydrate moietieson the PRO polypeptide is by chemical or enzymatic coupling ofglycosides to the polypeptide. Such methods are described in the art,e.g., in WO 87/05330 published Sep. 11, 1987, and in Aplin and Wriston,CRC Crit. Rev. Biochem., pp. 259-306 (1981).

[1309] Removal of carbohydrate moieties present on the PRO polypeptidemay be accomplished chemically or enzymatically or by mutationalsubstitution of codons encoding for amino acid residues that serve astargets for glycosylation. Chemical deglycosylation techniques are knownin the art and described, for instance, by Hakimuddin, et al., Arch.Biochem. Biophys., 259:52 (1987) and by Edge et al., Anal. Biochem.,118:131 (1981). Enzymatic cleavage of carbohydrate moieties onpolypeptides can be achieved by the use of a variety of endo- andexo-glycosidases as described by Thotakura et al., Meth. Enzymol.,138:350 (1987).

[1310] Another type of covalent modification of PRO comprises linkingthe PRO polypeptide to one of a variety of nonproteinaceous polymers,e.g., polyethylene glycol (PEG), polypropylene glycol, orpolyoxyalkylenes, in the manner set forth in U.S. Pat. No. 4,640,835;4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337.

[1311] The PRO of the present invention may also be modified in a way toform a chimeric molecule comprising PRO fused to another, heterologouspolypeptide or amino acid sequence.

[1312] In one embodiment, such a chimeric molecule comprises a fusion ofthe PRO with a tag polypeptide which provides an epitope to which ananti-tag antibody can selectively bind. The epitope tag is generallyplaced at the amino- or carboxyl- terminus of the PRO. The presence ofsuch epitope-tagged forms of the PRO can be detected using an antibodyagainst the tag polypeptide. Also, provision of the epitope tag enablesthe PRO to be readily purified by affinity purification using ananti-tag antibody or another type of affinity matrix that binds to theepitope tag. Various tag polypeptides and their respective antibodiesare well known in the art. Examples include poly-histidine (poly-his) orpoly-histidine-glycine (poly-his-gly) tags; the flu HA tag polypeptideand its antibody 12CA5 [Field et al., Mol. Cell. Biol., 8:2159-2165(1988)]; the c-myc tag and the 8F9, 3C7, 6E10, G4, B7 and 9E10antibodies thereto [Evan et al., Molecular and Cellular Biology,5:3610-3616 (1985)]; and the Herpes Simplex virus glycoprotein D (gD)tag and its antibody [Paborsky et al., Protein Engineering, (6):547-553(1990)]. Other tag polypeptides include the Flag-peptide [Hopp et al.,BioTechnologv, 6:1204-1210 (1988)]; the KT3 epitope peptide [Martin etal., Science, 255:192-194 (1992)]; an α-tubulin epitope peptide [Skinneret al., J. Biol. Chem., 266:15163-15166 (1991)]; and the 17 gene 10protein peptide tag [Lutz-Freyermuth et al., Proc. Natl. Acad. Sci. USA,87:6393-6397 (1990)].

[1313] In an alternative embodiment, the chimeric molecule may comprisea fusion of the PRO with an immunoglobulin or a particular region of animmunoglobulin. For a bivalent form of the chineric molecule (alsoreferred to as an “immunoadhesine”, such a fusion could be to the Fcregion of an IgG molecule. The Ig fusions preferably include thesubstitution of a soluble (transmembrane domain deleted or inactivated)form of a PRO polypeptide in place of at least one variable regionwithin an Ig molecule. In a particularly preferred embodiment, theimmunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge,CH1, CH2 and CH3 regions of an IgG1 molecule. For the production ofimmunoglobulin fusions see also U.S. Pat. No. 5,428,130 issued Jun. 27,1995.

[1314] D. Preparation of PRO

[1315] The description below relates primarily to production of PRO byculturing cells transformed or transfected with a vector containing PROnucleic acid. It is, of course, contemplated that alternative methods,which are well known in the art, may be employed to prepare PRO. Forinstance, the PRO sequence, or portions thereof, may be produced bydirect peptide synthesis using solid-phase techniques [see, e.g.,Stewart et al., Solid-Phase Peptide Synthesis, W.H. Freeman Co., SanFrancisco, Calif. (1969); Merrifield, J. Am. Chem. Soc., 85:2149-2154(1963)]. In vitro protein synthesis may be performed using manualtechniques or by automation. Automated synthesis may be accomplished,for instance, using an Applied Biosystems Peptide Synthesizer (FosterCity, Calif.) using manufacturer's instructions. Various portions of thePRO may be chemically synthesized separately and combined using chemicalor enzymatic methods to produce the full-length PRO.

[1316] 1. Isolation of DNA Encoding PRO

[1317] DNA encoding PRO may be obtained from a cDNA library preparedfrom tissue believed to possess the PRO mRNA and to express it at adetectable level. Accordingly, human PRO DNA can be convenientlyobtained from a cDNA library prepared from human tissue, such asdescribed in the Examples. The PRO- encoding gene may also be obtainedfrom a genomic library or by known synthetic procedures (e.g., automatednucleic acid synthesis).

[1318] Libraries can be screened with probes (such as antibodies to thePRO or oligonucleotides of at least about 20-80 bases) designed toidentify the gene of interest or the protein encoded by it. Screeningthe cDNA or genomic library with the selected probe may be conductedusing standard procedures, such as described in Sambrook et al.,Molecular Cloning: A Laboratory Manual (New York: Cold Spring HarborLaboratory Press, 1989). An alternative means to isolate the geneencoding PRO is to use PCR methodology [Sambrook et al., supra;Dieffenbach et al., PCR Primer: A Laboratory Manual (Cold Spring HarborLaboratory Press, 1995)].

[1319] The Examples below describe techniques for screening a cDNAlibrary. The oligonucleotide sequences selected as probes should be ofsufficient length and sufficiently unambiguous that false positives areminimized. The oligonucleotide is preferably labeled such that it can bedetected upon hybridization to DNA in the library being screened.Methods of labeling are well known in the art, and include the use ofradiolabels like ³²P-labeled ATP, biotinylation or enzyme labeling.Hybridization conditions, including moderate stringency and highstringency, are provided in Sambrook et al., supra.

[1320] Sequences identified in such library screening methods can becompared and aligned to other known sequences deposited and available inpublic databases such as GenBank or other private sequence databases.Sequence identity (at either the amino acid or nucleotide level) withindefined regions of the molecule or across the full-length sequence canbe determined using methods known in the art and as described herein.

[1321] Nucleic acid having protein coding sequence may be obtained byscreening selected cDNA or genomic libraries using the deduced aminoacid sequence disclosed herein for the first time, and, if necessary,using conventional primer extension procedures as described in Sambrooket al., supra, to detect precursors and processing intermediates of mRNAthat may not have been reverse-transcribed into cDNA.

[1322] 2. Selection and Transformation of Host Cells

[1323] Host cells are transfected or transformed with expression orcloning vectors described herein for PRO production and cultured inconventional nutrient media modified as appropriate for inducingpromoters, selecting transformants, or amplifying the genes encoding thedesired sequences. The culture conditions, such as media, temperature,pH and the like, can be selected by the skilled artisan without undueexperimentation. In general, principles, protocols, and practicaltechniques for maximizing the productivity of cell cultures can be foundin Mammalian Cell Biotechnology: a Practical Approach, M. Butler, ed.(IRL Press, 1991) and Sambrook et al., supra.

[1324] Methods of eukaryotic cell transfection and prokaryotic celltransformation are known to the ordinarily skilled artisan, for example,CaCl₂, CaPO₄, liposome-mediated and electroporation. Depending on thehost cell used, transformation is performed using standard techniquesappropriate to such cells. The calcium treatment employing calciumchloride, as described in Sambrook et al., supra, or electroporation isgenerally used for prokaryotes. Infection with Agrobactetium tumefaciensis used for transformation of certain plant cells, as described by Shawet al., Gene, 23:315 (1983) and WO 89/05859 published Jun. 29, 1989. Formammalian cells without such cell walls, the calcium phosphateprecipitation method of Graham and van der Eb, Virology, 52:456-457(1978) can be employed. General aspects of mammal cell host systemtransfections have been described in U.S. Pat. No. 4,399,216.Transformations into yeast are typically carried out according to themethod of Van Solingen et al., J. Bact., 130:946 (1977) and Hsiao etal., Proc. Natl. Acad. Sci. (USA), 76:3829 (1979). However, othermethods for introducing DNA into cells, such as by nuclearmicroinjection, electroporation, bacterial protoplast fusion with intactcells, or polycations, e.g., polybrene, polyornithine, may also be used.For various techniques for transforming mammalian cells, see Keown etal., Methods in Enzymology, 185:527-537 (1990) and Mansour et al.,Nature, 336:348-352 (1988).

[1325] Suitable host cells for cloning or expressing the DNA in thevectors herein include prokaryote, yeast, or higher eukaryote cells.Suitable prokaryotes include but are not limited to eubacteria, such asGram-negative or Gram-positive organisms, for example,Enterobacteriaceae such as E. coli. Various E. coli strains are publiclyavailable, such as E. coli K12 strain MM294 (ATCC 31,446); E. coil X1776(ATCC 31,537); E. coli strain W3110 (ATCC 27,325) and K5772 (ATCC53,635). Other suitable prokaryotic host cells includeEnterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter,Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium,Serratia, e.g., Serratia nmarcescans, and Shigella, as well as Bacillisuch as B. subtilis and B. licheniformis (e.g., B. licheniformis 41Pdisclosed in DD 266,710 published Apr. 12, 1989), Pseudomonas such as P.aeruginosa, and Streptomyces. These examples are illustrative ratherthan limiting. Strain W3110 is one particularly preferred host or parenthost because it is a commonhost strain for recombinant DNA productfermentations. Preferably, the host cell secretes minimal amounts ofproteolytic enzymes. For example, strain W3110 may be modified to effecta genetic mutation in the genes encoding proteins endogenous to thehost, with examples of such hosts including E. coli W3110 strain 1A2,which has the complete genotype tonA ; E. coli W3110 strain 9E4, whichhas the complete genotype tonA ptr3; E. coli W3110 strain 27C7 (ATCC55,244), which has the complete genotype tonA ptr3 phoA E15(argF-lac)169 degP ompT kan^(r) ; E. coil W3110 strain 37D6, which hasthe complete genotype tonA ptr3 phoA E15 (argF-lac)169 degP onipT rbs7ilvG kan^(r) ; E. coil W3110 strain 40B4, which is strain 37D6 with anon-kanamycin resistant degP deletion mutation; and an E. coli strainhaving mutant periplasmic protease disclosed in U.S. Pat. No. 4,946,783issued Aug. 7, 1990. Alternatively, in vitro methods of cloning, e.g.,PCR or other nucleic acid polymerase reactions, are suitable.

[1326] In addition to prokaryotes, eukaryotic microbes such asfilamentous fungi or yeast are suitable cloning or expression hosts forPRO-encoding vectors. Saccharomyces cerevisiae is a commonly used lowereukaryotic host microorganism. Others include Schizosaccharonzyces pombe(Beach and Nurse, Nature, 290:140 [1981]; EP 139,383 published May 2,1985); Kluyveromyces hosts (U.S. Pat. No. 4,943,529; Fleer et al.,Bio/Technology, 9:968-975(1991)) such as, e.g., K. lacis (MW98-8C,CBS683, CBS4574; Louvencourt et al., J. Bacteriol., 154(2):737-742[1983]), K. fragilis (ATCC 12,424), K. bulgaricas (ATCC 16,045), K.wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum(ATCC 36,906; Van den Berg et al., Bio/Technology, 8:135 (1990)), K.thernotolerans, and K. marxianus; yarrowia (EP 402,226); Pichia pastoris(EP 183,070; Sreekrishna et al., J. Basic Microbiol., 28:265-278[i988]); Candida; Trichoderma reesia (EP 244,234); Neurospora crassa(Case et al., Proc. Natl. Acad. Sci. USA, 76:5259-5263 [1979]);Schwannionyces such as Schwanniomyces occidentals (EP 394,538 publishedOct. 31, 1990); and filamentous fungi such as, e.g., Neurospora,Penicillium, Tolypocladium (WO 91/00357 published Jan. 10, 1991), andAspergillus hosts such as A. nidulans (Ballance et al., Biochem.Biophys. Res. Commun., 112:284-289 [1983]; Tilburn et al., Gene,26:205-221 [1983]; Yelton et al., Proc. Natl. Acad. Sci. USA, 81:1470-1474 [1984]) and A. niger (Kelly and Hynes, EMBO J., 4:475 -479[1985]). Methylotropic yeasts are suitable herein and include, but arenot limited to, yeast capable of growth on methanol selected from thegenera consisting of Hansenula, Candida, Kloeckera, Pichia,Saccharomyces, Torulopsis, and Rhodotorula. A list of specific speciesthat are exemplary of this class of yeasts may be found in C. Anthony,The Biochemistry of Methylotrophs, 269 (1982).

[1327] Suitable host cells for the expression of glycosylated PRO arederived from multicellular organisms. Examples of invertebrate cellsinclude insect cells such as Drosophila S2 and Spodoptera Sf9, as wellas plant cells. Examples of useful mammalian host cell lines includeChinese hamster ovary (CHO) and COS cells. More specific examplesinclude monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL1651); human embryonic kidney line (293 or 293 cells subdloned forgrowth in suspension culture, Graham et al., J. Gen Virol., 36:59(1977)); Chinese hamster ovary cells/-DHFR (CHO, Urlaub and Chasin,Proc. Natl. Acad. Sci. USA, 77:4216 (1980)); mouse sertoli cells (TM4,Mather, Biol. Reprod., 23:243-251 (1980)); human lung cells (W138, ATCCCCL 75); human liver cells (Hep G2, HB 8065); and mouse mammary tumor M060562, ATCC CCL51). The selection of the appropriate host cell isdeemed to be within the skill in the art.

[1328] 3. Selection and Use of a Replicable Vector

[1329] The nucleic acid (e.g., cDNA or genomic DNA) encoding PRO may beinserted into a replicable vector for cloning (amplification of the DNA)or for expression. Various vectors are publicly available. The vectormay, for example, be in the form of a plasmid, cosmid, viral particle,or phage. The appropriate nucleic acid sequence may be inserted into thevector by a variety of procedures. In general, DNA is inserted into anappropriate restriction endonuclease site(s) using techniques known inthe art. Vector components generally include, but are not limited to,one or more of a signal sequence, an origin of replication, one or moremarker genes, an enhancer element, a promoter, and a transcriptiontermination sequence. Construction of suitable vectors containing one ormore of these components employs standard ligation techniques which areknown to the skilled artisan.

[1330] The PRO may be produced recombinantly not only directly, but alsoas a fusion polypeptide with a heterologous polypeptide, which may be asignal sequence or other polypeptide having a specific cleavage site atthe N-terminus of the mature protein or polypeptide. In general, thesignal sequence may be a component of the vector, or it may be a part ofthe PRO-encoding DNA that is inserted into the vector. The signalsequence may be a prokaryotic signal sequence selected, for example,from the group of the alkaline phosphatase, penicillinase, lpp, orheat-stable enterotoxin II leaders. For yeast secretion the signalsequence may be, e.g., the yeast invertase leader, alpha factor leader(including Saccharomyces and Kluyveroffyces α-factor leaders, the latterdescribed in U.S. Pat. No. 5,010,182), or acid phosphatase leader, theC. albicans glucoamylase leader (EP 362,179 published Apr. 4, 1990), orthe signal described in WO 90113646 published Nov. 15, 1990. Inmammalian cell expression, mammalian signal sequences may be used todirect secretion of the protein, such as signal sequences from secretedpolypeptides of the same or related species, as well as viral secretoryleaders.

[1331] Both expression and cloning vectors contain a nucleic acidsequence that enables the vector to replicate in one or more selectedhost cells. Such sequences are well known for a variety of bacteria,yeast, and viruses. The origin of replication from the plasmid pBR322 issuitable for most Gram-negative bacteria, the 21a plasmid origin issuitable for yeast, and various viral origins (SV40, polyoma,adenovirus, VSV or BPV) are useful for cloning vectors in mammaliancells.

[1332] Expression and cloning vectors will typically contain a selectiongene, also termed a selectable marker. Typical selection genes encodeproteins that (a) confer resistance to antibiotics or other toxins,e.g., ampicilln, neomycin, methotrexate, or tetracycline, (b) complementauxotrophic deficiencies, or (c) supply critical nutrients not availablefrom complex media, e.g., the gene encoding D-alanine racemase forBacilli.

[1333] An example of suitable selectable markers for mammalian cells arethose that enable the identification of cells competent to take up thePRO-encoding nucleic acid, such as DHFR or thymidine kinase. Anappropriate host cell when wild-type DHFR is employed is the CHO cellline deficient in DHFR activity, prepared and propagated as described byUrlaub et al., Proc. Natl. Acad. Sci. USA, 77:4216 (1980). A suitableselection gene for use in yeast is the trp1 gene present in the yeastplasmid YRp7 [Stinchcomb et al., Nature, 282:39 (1979); Kingsman et al.,Gene, 7:141 (1979); Tschemper et al., Gene, 10:157 (1980)]. The trp1gene provides a selection marker for a mutant strain of yeast lackingthe ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1[Jones, Genetics, 85:12 (1977)].

[1334] Expression and cloning vectors usually contain a promoteroperably linked to the PROncoding nucleic acid sequence to direct mRNAsynthesis. Promoters recognized by a variety of potential host cells arewell known. Promoters suitable for use with prokaryotic hosts includethe β-lactamase and lactose promoter systems [Chang et al., Nature,275:615 (1978); Goeddel et al., Nature, 281:544 (1979)], alkalinephosphatase, a tryptophan (trp) promoter system [Goeddel, Nucleic AcidsRes., 8:4057 (1980); EP 36,776], and hybrid promoters such as the tacpromoter [deBoer et al., Proc. Natl. Acad. Sci. USA, 80:21-25 (1983)].Promoters for use in bacterial systems also will contain aShine-Dalgarno (S.D.) sequence operably linked to the DNA encoding PRO.

[1335] Examples of suitable promoting sequences for use with yeast hostsinclude the promoters for 3-phosphoglycerate kinase [Hitzeman et al., J.Biol. Chem., 255:2073 (1980)] or other glycolytic enzymes [Hess et al.,J. Adv. Enzyme Reg., 7:149 (1968); Holland, Biochemistry, 17:4900(1978)], such as enolase, glyceraldehyde-3-phosphate dehydrogenase,hexokiuase, pyruvate decarboxylase, phosphofructokinase,glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvatekinase, triosephosphate isomerase, phosphoglucose isomerase, andglucokinase.

[1336] Other yeast promoters, which are inducible promoters having theadditional advantage of transcription controlled by growth conditions,are the promoter regions for alcohol dehydrogenase 2, isocytochrome C,acid phosphatase, degradative enzymes associated with nitrogenmetabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase,and enzymes responsible for maltose and galactose utilization. Suitablevectors and promoters for use in yeast expression are further describedin EP 73,657.

[1337] PRO transcription from vectors in mammalian host cells iscontrolled, for example, by promoters obtained from the genomes ofviruses such as polyoma virus, fowlpox virus (UK 2,211,504 publishedJul. 5, 1989), adenovirus (such as Adenovirus 2), bovine papillomavirus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-Bvirus and Simian Virus 40 (SV40), from heterologous mammalian promoters,e.g., the actin promoter or an immunoglobulin promoter, and fromheat-shock promoters, provided such promoters are compatible with thehost cell systems.

[1338] Transcription of a DNA encoding the PRO by higher eukaryotes maybe increased by inserting an enhancer sequence into the vector.Enhancers are cis-acting elements of DNA, usually about from 10 to 300bp, that act on a promoter to increase its transcription. Many enhancersequences are now known from mammalian genes (globin, elastase, albumin,c-fetoprotein, and insulin). Typically, however, one will use anenhancer from a eukaryotic cell virus. Examples include the SV40enhancer on the late side of the replication origin (bp 100-270), thecytomegalovirus early promoter enhancer, the polyoma enhancer on thelate side of the replication origin, and adenovirus enhancers. Theenhancer may be spliced into the vector at a position 5′ or 3′ to thePRO coding sequence, but is preferably located at a site 5′ from thepromoter.

[1339] Expression vectors used ineukaryotic host cells (yeast, fungi,insect, plant, animal, human, or nucleated cells from othermulticellular organisms) will also contain sequences necessary for thetermination of transcription and for stabilizing the mRNA. Suchsequences are commonly available from the 5′ and, occasionally 3′,untranslated regions of eukaryotic or viral DNAs or cDNAs. These regionscontain nucleotide segments transcribed as polyadenylated fragments inthe untranslated portion of the mRNA encoding PRO.

[1340] Still other methods, vectors, and host cells suitable foradaptation to the synthesis of PRO in recombinant vertebrate cellculture are described in Gething et al., Nature, 293:620-625 (1981);Mantei et al., Nature, 281:4046 (1979); EP 117,060; and EP 117,058.

[1341] 4. Detecting Gene Amplification/Expression

[1342] Gene amplification and/or expression may be measured in a sampledirectly, for example, by conventional Southern blotting, Northernblotting to quantitate the transcription of mRNA [Thomas, Proc. Natl.Acad. Sci. USA, 77:5201-5205 (1980)], dot blotting (DNA analysis), or insitu hybridization, using an appropriately labeled probe, based on thesequences provided herein. Alternatively, antibodies may be employedthat can recognize specific duplexes, including DNA duplexes, RNAduplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes. Theantibodies in turn may be labeled and the assay may be carried out wherethe duplex is bound to a surface, so that upon the formation of duplexon the surface, the presence of antibody bound to the duplex can bedetected.

[1343] Gene expression, alternatively, may be measured by immunologicalmethods, such as immunohistochemical staining of cells or tissuesections and assay of cell culture or body fluids, to quantitatedirectly the expression of gene product. Antibodies useful forimmunohistochemical staining and/or assay of sample fluids may be eithermonoclonal or polyclonal, and may be prepared in any mammal.Conveniently, the antibodies may be prepared against a native sequencePRO polypeptide or against a synthetic peptide based on the DNAsequences provided herein or against exogenous sequence fused to PRO DNAand encoding a specific antibody epitope.

[1344] 5. Purification of Polypeptide

[1345] Forms of PRO may be recovered from culture medium or from hostcell lysates. If membrane-bound, it can be released from the membraneusing a suitable detergent solution (e.g. Triton-X 100) or by enzymaticcleavage. Cells employed in expression of PRO can be disrupted byvarious physical or chemical means, such as freeze-thaw cycling,sonication, mechanical disruption, or cell lysing agents.

[1346] It may be desired to purify PRO from recombinant cell proteins orpolypeptides. The following procedures are exemplary of suitablepurification procedures: by fractionation on an ion-exchange column;ethanol precipitation; reverse phase HPLC; chromatography on silica oron a cation-exchange resin such as DEAE; chromatofocusmg; SDS-PAGE;ammonium sulfate precipitation; gel filtration using, for example,Sephadex G-75; protein A Sepharose columns to remove contaminants suchas IgG; and metal chelating columns to bind epitope-tagged forms of thePRO. Various methods of protein purification may be employed and suchmethods are known in the art and described for example in Deutscher,Methods in Enzymology, 182 (1990); Scopes, Protein Purification:Principles and Practice, Springer-Verlag, New York (1982). Thepurification step(s) selected will depend, for example, on the nature ofthe production process used and the particular PRO produced.

[1347] E. Uses for PRO

[1348] Nucleotide sequences (or their complement) encoding PRO havevarious applications in the art of molecular biology, including uses ashybridization probes, in chromosome and gene mapping and in thegeneration of anti-sense RNA and DNA. PRO nucleic acid will also beuseful for the preparation of PRO polypeptides by the recombinanttechniques described herein.

[1349] The full-Length native sequence PRO gene, or portions thereof,may be used as hybridization probes for a cDNA library to isolate thefull-length PRO cDNA or to isolate still other cDNAs (for instance,those encoding naturally-occurring variants of PRO or PRO from otherspecies) which have a desired sequence identity to the native PROsequence disclosed herein. Optionally, the length of the probes will beabout 20 to about 50 bases. The hybridization probes may be derived fromat least partially novel regions of the full length native nucleotidesequence wherein those regions may be determined without undueexperimentation or from genomic sequences including promoters, enhancerelements and introns of native sequence PRO. By way of example, ascreening method will comprise isolating the coding region of the PROgene using the known DNA sequence to synthesize a selected probe ofabout 40 bases. Hybridization probes may be labeled by a variety oflabels, including radionucleotides such as ³²P or 35S, or enzymaticlabels such as alkaline phosphatase coupled to the probe viaavidin/biotin coupling systems. Labeled probes having a sequencecomplementary to that of the PRO gene of the present invention can beused to screen libraries of human cDNA, genomic DNA or mRNA to determinewhich members of such libraries the probe hybridizes to. Hybridizationtechniques are described in further detail in the Examples below.

[1350] Any EST sequences disclosed in the present application maysimilarly be employed as probes, using the methods disclosed herein.

[1351] Other useful fragments of the PRO nucleic acids include antisenseor sense oligonucleotides comprising a singe-stranded nucleic acidsequence (either RNA or DNA) capable of binding to target PRO mRNA(sense) or PRO DNA (antisense) sequences. Antisense or senseoligonucleotides, according to the present invention, comprise afragment of the coding region of PRO DNA. Such a fragment generallycomprises at least about 14 nucleotides, preferably from about 14 to 30nucleotides. The ability to derive an antisense or a senseoligonucleotide, based upon a cDNA sequence encoding a given protein isdescribed in, for example, Stein and Cohen (Cancer Res. 48:2659, 1988)and van der Krol et al. (BioTechniques 6:958, 1988).

[1352] Binding of antisense or sense oligonucleotides to target nucleicacid sequences results in the formation of duplexes that blocktranscription or translation of the target sequence by one of severalmeans, including enhanced degradation of the duplexes, prematuretermination of transcription or translation, or by other means. Theantisense oligonucleotides thus may be used to block expression of PROproteins. Antisense or sense oligonucleotides further compriseoligonucleotides having modified sugar-phosphodiester backbones (orother sugar linkages, such as those described in WO 91/06629) andwherein such sugar linkages are resistant to endogenous nucleases. Sucholigonucleotides with resistant sugar linkages are stable in vivo (i.e.,capable of resisting enzymatic degradation) but retain sequencespecificity to be able to bind to target nucleotide sequences.

[1353] Other examples of sense or antisense oligonucleotides includethose oligonucleotides which are covalently linked to organic moieties,such as those described in WO 90/10048, and other moieties thatincreases affinity of the oligonucleotide for a target nucleic acidsequence, such as poly-(L-lysine). Further still, intercalating agents,such as ellipticine, and alkylating agents or metal complexes may beattached to sense or antisense oligonucleotides to modify bindingspecificities of the antisense or sense oligonucleotide for the targetnucleotide sequence.

[1354] Antisense or sense oligonucleotides may be introduced into a cellcontaining the target nucleic acid sequence by any gene transfer method,including, for example, CaPO₄-mediated DNA transfection,electroporation, or by using gene transfer vectors such as Epstein-Barrvirus. In a preferred procedure, an antisense or sense oligonucleotideis inserted into a suitable retroviral vector. A cell containing thetarget nucleic acid sequence is contacted with the recombinantretroviral vector, either in vivo or ex vivo. Suitable retroviralvectors include, but are not limited to, those derived from the murineretrovirus M-MuLV, N2 (a retrovirus derived from M-MuLV), or the doublecopy vectors designated DCT5A, DCT5B and DCTSC (see WO 90/13641).

[1355] Sense or antisense oligonucleotides also may be introduced into acell containing the target nucleotide sequence by formation of aconjugate with a ligand binding molecule, as described in WO 91/04753.Suitable ligand binding molecules include, but are not limited to, cellsurface receptors, growth factors, other cytokines, or other ligandsthat bind to cell surface receptors. Preferably, conjugation of theligand binding molecule does not substantially interfere with theability of the ligand binding molecule to bind to its correspondingmolecule or receptor, or block entry of the sense or antisenseoligonucleotide or its conjugated version into the cell.

[1356] Alternatively, a sense or an antisense oligonucleotide may beintroduced into a cell containing the target nucleic acid sequence byformation of an oligonucleotide-lipid complex, as described in WO90110448. The sense or antisense oligonucleotide-lipid complex ispreferably dissociated within the cell by an endogenous lipase.

[1357] Antisense or sense RNA or DNA molecules are generally at leastabout 5 bases in length, about 10 bases in length, about 15 bases inlength, about 20 bases in length, about 25 bases in length, about 30bases in length, about 35 bases in length, about 40 bases in length,about 45 bases in length, about 50 bases in length, about 55 bases inlength, about 60 bases in length, about 65 bases in length, about 70bases in length, about 75 bases in length, about 80 bases in length,about 85 bases in length, about 90 bases in length, about 95 bases inlength, about 100 bases in length, or more.

[1358] The probes may also be employed in PCR techniques to generate apool of sequences for identification of closely related PRO codingsequences.

[1359] Nucleotide sequences encoding a PRO can also be used to constructhybridization probes for mapping the gene which encodes that PRO and forthe genetic analysis of individuals with genetic disorders. Thenucleotide sequences provided herein may be mapped to a chromosome andspecific regions of a chromosome using known techniques, such as in situhybridization, linkage analysis against known chromosomal markers, andhybridization screening with libraries.

[1360] When the coding sequences for PRO encode a protein which binds toanother protein (example, where the PRO is a receptor), the PRO can beused in assays to identify the other proteins or molecules involved inthe binding interaction. By such methods, inhibitors of thereceptor/ligand binding interaction can be identified. Proteins involvedin such binding interactions can also be used to screen for peptide orsmall molecule inhibitors or agonists of the binding interaction. Also,the receptor PRO can be used to isolate correlative ligand(s). Screeningassays can be designed to find lead compounds that mimic the biologicalactivity of a native PRO or a receptor for PRO. Such screening assayswill include assays amenable to high-throughput screening of chemicallibraries, making them particularly suitable for identifying smallmolecule drug candidates. Small molecules contemplated include syntheticorganic or inorganic compounds. The assays can be performed in a varietyof formats, including protein-protein binding assays, biochemicalscreening assays, immunoassays and cell based assays, which are wellcharacterized in the art.

[1361] Nucleic acids which encode PRO or its modified forms can also beused to generate either transgenic animals or “knock out” animals which,in turn, are useful in the development and screening of therapeuticallyuseful reagents. A transgenic animal (e.g., a mouse or rat) is an animalhaving cells that contain a transgene, which transgene was introducedinto the animal or an ancestor of the animal at a prenatal, e.g., anembryonic stage. A transgene is a DNA which is integrated into thegenome of a cell from which a transgenic animal develops. In oneembodiment, cDNA encoding PRO can be used to clone genomic DNA encodingPRO in accordance with established techniques and the genomic sequencesused to generate transgenic animals that contain cells which express DNAencoding PRO. Methods for generating transgenic animals, particularlyanimals such as mice or rats, have become conventional in the art andare described, for example, in U.S. Pat. Nos. 4,736,866 and 4,870,009.Typically, particular cells would be targeted for PRO transgeneincorporation with tissue-specific enhancers. Transgenic animals thatinclude a copy of a transgene encoding PRO introduced into the germ lineof the animal at an embryonic stage can be used to examine the effect ofincreased expression of DNA encoding PRO. Such animals can be used astester animals for reagents thought to confer protection from, forexample, pathological conditions associated with its overexpression. Inaccordance with this facet of the invention, an animal is treated withthe reagent and a reduced incidence of the pathological condition,compared to untreated animals bearing the transgene, would indicate apotential therapeutic intervention for the pathological condition.

[1362] Alternatively, non-human homologues of PRO can be used toconstruct a PRO “knock out” animal .which has a defective or alteredgene encoding PRO as a result of homologous recombination between theendogenous gene encoding PRO and altered genomic DNA encoding PROintroduced into an embryonic stem cell of the animal. For example, cDNAencoding PRO can be used to clone genomic DNA encoding PRO in accordancewith established techniques. A portion of the genomic DNA encoding PROcan be deleted or replaced with another gene, such as a gene encoding aselectable marker which can be used to monitor integration. Typically,several kilobases of unaltered flanking DNA (both at the 5′ and 3′ ends)are included in the vector [see e.g., Thomas and Capeochi, Cell, 51:503(1987) for a description of homologous recombination vectors]. Thevector is introduced into an embryonic stem cell line (e.g., byelectroporation) and cells in which the introduced DNA has homologouslyrecombined with the endogenous DNA are selected [see e.g., Li et al.,Cell, 69:915 (1992)]. The selected cells are then injected into ablastocyst of an animal (e.g., a mouse or rat) to form aggregationchimeras [see e.g., Bradley, in Teratocarcinomas and Embryonic StemCells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987),pp. 113-152]. A chimeric embryo can then be implanted into a suitablepseudopregnant female foster animal and the embryo brought to term tocreate a “knock out” animal. Progeny harboring the homologouslyrecombined DNA in their germ cells can be identified by standardtechniques and used to breed animals in which all cells of the animalcontain the homologously recombined DNA. Knockout animals canbecharacterized for instance, for their ability to defend against certainpathological conditions and for their development of pathologicalconditions due to absence of the PRO polypeptide.

[1363] Nucleic acid encoding the PRO polypeptides may also be used ingene therapy. In gene therapy applications, genes are introduced intocells in order to achieve in vivo synthesis of a therapeuticallyeffective genetic product, for example for replacement of a defectivegene. “Gene therapy” includes both conventional gene therapy where alasting effect is achieved by a single treatment, and the administrationof gene therapeutic agents, which involves the one time or repeatedadministration of a therapeutically effective DNA or mRNA. AntisenseRNAs and DNAs can be used as therapeutic agents for blocking theexpression of certain genes in vivo. It has already been shown thatshort antisense oligonucleotides can be imported into cells where theyact as inhibitors, despite their low intracellular concentrations causedby their restricted uptake by the cell membrane. (Zamecnik et al., Proc.Natl. Acad. Sci. USA 83:4143-4146 [1986]). The oligonucleotides can bemodified to enhance their uptake, e.g. by substituting their negativelycharged phosphodiester groups by uncharged groups.

[1364] There are a variety of techniques available for introducingnucleic acids into viable cells. The techniques vary depending uponwhether the nucleic acid is transferred into cultured cells in vitro, orin vivo in the cells of the intended host. Techniques suitable for thetransfer of nucleic acid into mammalian cells in vitro include the useof liposomes, electroporation, microinjection, cell fusion,DEAE-dextran, the calcium phosphate precipitation method, etc. Thecurrently preferred in vivo gene transfer techniques includetransfection with viral (typically retroviral) vectors and viral coatprotein-liposome mediated transfection (Dzau et al., Trends inBiotechnology 11, 205-210 [1993]). In some situations it is desirable toprovide the nucleic acid source with an agent that targets the targetcells, such as an antibody specific for a cell surface membrane proteinor the target cell, a ligand for a receptor on the target cell, etc.Where liposomes are employed, proteins which bind to a cell surfacemembrane protein associated with endocytosis may be used for targetingand/or to facilitate uptake, e.g. capsid proteins or fragments thereoftropic for a particular cell type, antibodies for proteins which undergointernalization in cycling, proteins that target intracellularlocalization and enhance intracellular half-life. The technique ofreceptor-mediated endocytosis is described, for example, by Wu et al.,J. Biol. Chem. 262, 4429-4432 (1987); and Wagner et al., Proc. Natl.Acad. Sci. USA 87, 3410-3414 (1990). For review of gene marking and genetherapy protocols see Anderson et al., Science 256, 808-813 (1992).

[1365] The PRO polypeptides described herein may also be employed asmolecular weight markers for protein electrophoresis purposes and theisolated nucleic acid sequences may be used for recombinantly expressingthose markers.

[1366] The nucleic acid molecules encoding the PRO polypeptides orfragments thereof described herein are useful for chromosomeidentification. In this regard, there exists an ongoing need to identifynew chromosome markers, since relatively few chromosome markingreagents, based upon actual sequence data are presently available. EachPRO nucleic acid molecule of the present invention can be used as achromosome marker.

[1367] The PRO polypeptides and nucleic acid molecules of the presentinvention may also be used for tissue typing, wherein the PROpolypeptides of the present invention may be differentially expressed inone tissue as compared to another. PRO nucleic acid molecules will finduse for generating probes for PCR, Northern analysis, Southern analysisand Western analysis.

[1368] The PRO polypeptides described herein may also be employed astherapeutic agents. The PRO polypeptides of the present invention can beformulated according to known methods to prepare pharmaceutically usefulcompositions, whereby the PRO product hereof is combined in admixturewith a pharmaceutically acceptable carrier vehicle. Therapeuticformulations are prepared for storage by mixing the active ingredienthaving the desired degree of purity with optional physiologicallyacceptable carriers, excipients or stabilizers (Remington'sPharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the formof lyophilized formulations or aqueous solutions. Acceptable carriers,excipients or stabilizers are nontoxic to recipients at the dosages andconcentrations employed, and include buffers such as phosphate, citrateand other organic acids; antioxidants including ascorbic acid; lowmolecular weight (less than about 10 residues) polypeptides; proteins,such as serum albumin, gelatin or imnunoglobulins; hydrophilic polymerssuch as polyvinylpyrrolidone, amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides and othercarbohydrates including glucose, mannose, or dextrins; chelating agentssuch as EDTA; sugar alcohols such as mannitol or sorbitol; salt-formingcounterions such as sodium; and/or nonionic surfactints such as TWEEN™,PLURONICS™ or PEG.

[1369] The formulations to be used for in vivo administration must besterile. This is readily accomplished by filtration through sterilefiltration membranes, prior to or following lyophilization andreconstitution.

[1370] Therapeutic compositions herein generally are placed into acontainer having a sterile access port, for example, an intravenoussolution bag or vial having a stopper pierceable by a hypodermicinjection needle.

[1371] The route of administration is in accord with known methods, e.g.injection or infusion by intravenous, intraperitoneal, intracerebral,intramuscular, intraocular, intraarterial or intralesional routes,topical administration, or by sustained release systems.

[1372] Dosages and desired drug concentrations of pharmaceuticalcompositions of the present invention may vary depending on theparticular use envisioned. The determination of the appropriate dosageor route of administration is well within the skill of an ordinaryphysician. Animal experiments provide reliable guidance for thedetermination of effective doses for human therapy. Interspecies scalingof effective doses can be performed following the principles laid downby Mordenti, J. and Chappell, W. “The use of interspecies scaling intoxicokinetics” In Toxicokinetics and New Drug Development, Yacobi etal., Eds., Pergamon Press, New York 1989, pp. 42-96.

[1373] When in vivo administration of a PRO polypeptide or agonist orantagonist thereof is employed, normal dosage amounts may vary fromabout 10 ng/kg to up to 100 mg/kg of mammal body weight or more per day,preferably about 1 μg/kg/day to 10 mg/kg/day, depending upon the routeof administration. Guidance as to particular dosages and methods ofdelivery is provided in the literature; see, for example, U.S. Pat. Nos.4,657,760; 5,206,344; or 5,225,212. It is anticipated that differentformulations will be effective for different treatment compounds anddifferent disorders, that administration targeting one organ or tissue,for example, may necessitate delivery in a manner different from that toanother organ or tissue.

[1374] Where sustained-release administration of a PRO polypeptide isdesired in a formulation with release characteristics suitable for thetreatment of any disease or disorder requiring administration of the PROpolypeptide, microencapsulation of the PRO polypeptide is contemplated.Microencapsulation of recombinant proteins for sustained release hasbeen successfully performed with human growth hormone (rhGH),interferon-(rhIFN−), interleuldn-2, and MN rgpl20. Johnson et al., Nat.Med., 2:795-799 (1996); Yasuda, Biomed. Ther., 27:1221-1223 (1993); Horaet al., Bio/Technology, 8:755-758 (1990); Cleland, “Design andProduction of Single Imunization Vaccines Using PolylactidePolyglycolide Microsphere Systems,” in Vaccine Design: The Subunit andAdjuvant Aproach, Powell and Newman, eds, (Plenum Press: New York,1995), pp. 439-462; WO 97/03692, WO 96/40072, WO 96107399; and U.S. Pat.No. 5,654,010.

[1375] The sustained-release formulations of these proteins weredeveloped using poly-lactic-coglycolic acid (PLGA) polymer due to itsbiocompatibility and wide range of biodegradable properties. Thedegradation products of PLGA, lactic and glycolic acids, can be clearedquickly within the human body. Moreover, the degradability of thispolymer can be adjusted from months to years depending on its molecularweight and composition. Lewis, “Controlled release of bioactive agentsfrom lactide/glycolide polymer,” in: M. Chasin and R. Langer (Eds.),Biodegradable Polymers as Drug Delivery Systems (Marcel Dekker: NewYork, 1990), pp. 141.

[1376] This invention encompasses methods of screening compounds toidentify those that mimic the PRO polypeptide (agonists) or prevent theeffect of the PRO polypeptide (antagonists). Screening assays forantagonist drug candidates are designed to identify compounds that bindor complex with the PRO polypeptides encoded by the genes identifiedherein, or otherwise interfere with the interaction of the encodedpolypeptides with other cellular proteins. Such screening assays willinclude assays amenable to high-throughput screening of chemicallibraries, making them particularly suitable for identifying smallmolecule drug candidates.

[1377] The assays can be performed in a variety of formats, includingprotein-protein binding assays, biochemical screening assays,immunoassays, and cell-based assays, which are well characterized in theart.

[1378] All assays for antagonists are common in that they call forcontacting the drug candidate with a PRO polypeptide encoded by anucleic acid identified herein under conditions and for a timesufficient to allow these two components to interact.

[1379] In binding assays, the interaction is binding and the complexformed can be isolated or detected in the reaction mixture. In aparticular embodiment, the PRO polypeptide encoded by the geneidentified herein or the drug candidate is immobilized on a solid phase,e.g., on a microtiter plate, by covalent or non-covalent attachments.Non-covalent attachment generally is accomplished by coating the solidsurface with a solution of the PRO polypeptide and drying.Alternatively, an immobilized antibody, e.g., a monoclonal antibody,specific for the PRO polypeptide to be immobilized can be used to anchorit to a solid surface. The assay is performed by adding thenon-immobilized component, which may be labeled by a detectable label,to the immobilized component, e.g., the coated surface containing theanchored component. When the reaction is complete, the non-reactedcomponents are removed, e.g., by washing, and complexes anchored on thesolid surface are detected. When the originally non-immobilizedcomponent carries a detectable label, the detection of label immobilizedon the surface indicates that complexing occurred. Where the originallynon-immobilized component does not carry a label, complexing can bedetected, for example, by using a labeled antibody specifically bindingthe immobilized complex.

[1380] If the candidate compound interacts with but does not bind to aparticular PRO polypeptide encoded by a gene identified herein, itsinteraction with that polypeptide can be assayed by methods well knownfor detecting protein-protein interactions. Such assays includetraditional approaches, such as, e.g., cross-linking, co-immunoprecipitation, and co-purification through gradients orchromatographic columns. In addition, protein- protein interactions canbe monitored by using a yeast-based genetic system described by Fieldsand co-workers (Fields and Song, Nature (London), 340:245-246 (1989);Chien et al., Proc. Natl. Acad. Sci. USA, 88:9578-9582 (1991)) asdisclosed by Chevray and Nathans, Proc. Natl. Acad. Sci. USA, 89:5789-5793 (1991). Many transcriptional activators, such as yeast GAL4,consist of two physically discrete modular domains, one acting as theDNA-binding domain, the other one functioning as thetranscription-activation domain. The yeast expression system describedin the foregoing publications (generally referred to as the “two-hybridsystem”) takes advantage of this property, and employs two hybridproteins, one in which the target protein is fused to the DNA-bindingdomain of GAL4, and another, in which candidate activating proteins arefused to the activation domain. The expression of a GAL1-lacZ reportergene under control of a GAL4-activated promoter depends onreconstitution of GAL4 activity via protein-protein interaction.Colonies containing interacting polypeptides are detected with achromogenic substrate for β-galactosidase. A complete kit (MATCHMAKER™)for identifying protein-protein interactions between two specificproteins using the two- hybrid technique is commercially available fromClontech. This system can also be extended to map protein domainsinvolved in specific protein interactions as well as to pinpoint aminoacid residues that are crucial for these interactions.

[1381] Compounds that interfere with the interaction of a gene encodinga PRO polypeptide identified herein and other intra- or extracellularcomponents can be tested as follows: usually a reaction mixture isprepared containing the product of the gene and the intra- orextracellular component under conditions and for a time allowing for theinteraction and binding of the two products. To test the ability of acandidate compound to inhibit binding, the reaction is run in theabsence and in the presence of the test compound. In addition, a placebomay be added to a third reaction mixture, to serve as positive control.The binding (complex formation) between the test compound and the intra-or extracellular component present in the mixture is monitored asdescribed hereinabove. The formation of a complex in the controlreaction(s) but not in the reaction mixture containing the test compoundindicates that the test compound interferes with the interaction of thetest compound and its reaction partner.

[1382] To assay for antagonists, the PRO polypeptide may be added to acell along with the compound to be screened for a particular activityand the ability of the compound to inhibit the activity of interest inthe presence of the PRO polypeptide indicates that the compound is anantagonist to the PRO polypeptide. Alternatively, antagonists may bedetected by combining the PRO polypeptide and a potential antagonistwith membrane-bound PRO polypeptide receptors or recombinant receptorsunder appropriate conditions for a competitive inhibition assay. The PROpolypeptide can be labeled, such as by radioactivity, such that thenumber of PRO polypeptide molecules bound to the receptor can be used todetermine the effectiveness of the potential antagonist. The geneencoding the receptor can be identified by numerous methods known tothose of skill in the art, for example, ligand panring and FACS sorting.Coligan et al., Current Protocols in Immun., 1(2): Chapter 5 (1991).Preferably, expression cloning is employed wherein polyadenylated RNA isprepared from a cell responsive to the PRO polypeptide and a cDNAlibrary created from this RNA is divided into pools and used totransfect COS cells or other cells that are not responsive to the PROpolypeptide. Transfected cells that are grown on glass slides areexposed to labeled PRO polypeptide. The PRO polypeptide can be labeledby a variety of means including iodination or inclusion of a recognitionsite for a site-specific protein kinase. Following fixation andincubation, the slides are subjected to autoradiographic analysis.Positive pools are identified and sub-pools are prepared andre-transfected using an interactive sub-pooling and re-screeningprocess, eventually yielding a single clone that encodes the putativereceptor.

[1383] As an alternative approach for receptor identification, labeledPRO polypeptide can be photoaffinity-linked with cell membrane orextract preparations that express the receptor molecule. Cross-linkedmaterial is resolved by PAGE and exposed to X-ray film. The labeledcomplex containing the receptor can be excised, resolved into peptidefragments, and subjected to protein micro-sequencing. The amino acidsequence obtained from micro- sequencing would be used to design a setof degenerate oligonucleotide probes to screen a cDNA library toidentify the gene encoding the putative receptor.

[1384] In another assay for antagonists, mammalian cells or a membranepreparation expressing the receptor would be incubated with labeled PROpolypeptide in the presence of the candidate compound. The ability ofthe compound to enhance or block this interaction could then bemeasured.

[1385] More specific examples of potential antagonists include anoligonucleotide that binds to the fusions of immunoglobulin with PROpolypeptide, and, in particular, antibodies including, withoutlimitation, poly- and monoclonal antibodies and antibody fragments,single-chain antibodies, anti-idiotypic antibodies, and chimeric orhumanized versions of such antibodies or fragments, as well as humanantibodies and antibody fragments. Alternatively, a potential antagonistmay be a closely related protein, for example, a mutated form of the PROpolypeptide that recognizes the receptor but imparts no effect, therebycompetitively inhibiting the action of the PRO polypeptide.

[1386] Another potential PRO polypeptide antagonist is an antisense RNAor DNA construct prepared using antisense technology, where, e.g., anantisense RNA or DNA molecule acts to block directly the translation ofmRNA by hybridizing to targeted mRNA and preventing protein translation.Antisense technology can be used to control gene expression throughtriple-helix formation or antisense DNA or RNA, both of which methodsare based on binding of a polynucleotide to DNA or RNA. For example, the5′ coding portion of the polynucleotide sequence, which encodes themature PRO polypeptides herein, is used to design an antisense RNAoligonucleotide of from about 10 to 40 base pairs in length. A DNAoligonucleotide is designed to be complementary to a region of the geneinvolved in transcription (triple helix—see Lee et al., Nucl. AcidsRes., 6:3073 (1979); Cooney et al., Science, 241: 456 (1988); Dervan etal., Science, 251:1360 (1991)), thereby preventing transcription and theproduction of the PRO polypeptide. The antisense RNA oligonucleotidehybridizes to the mRNA in vivo and blocks translation of the mRNAmolecule into the PRO polypeptide (antisense—Okano, Neurochem., 56:560(1991); Oligodeoxynucleotides as Antisense inhibitors of Gene Expression(CRC Press: Boca Raton, Fla., 1988). The oligonucleotides describedabove can also be delivered to cells such that the antisense RNA or DNAmay be expressed in vivo to inhibit production of the PRO polypeptide.When antisense DNA is used, oligodeoxyribonucleotides derived from thetranslation-initiation site, e.g., between about −10 and +10 positionsof the target gene nucleotide sequence, are preferred.

[1387] Potential antagonists include small molecules that bind to theactive site, the receptor binding site, or growth factor or otherrelevant binding site of the PRO polypeptide, thereby blocking thenormal biological activity of the PRO polypeptide. Examples of smallmolecules include, but are not limited to, small peptides orpeptide-like molecules, preferably soluble peptides, and syntheticnon-peptidyl organic or inorganic compounds.

[1388] Ribozymes are enzymatic RNA molecules capable of catalyzing thespecific cleavage of RNA. Ribozymes act by sequence-specifichybridization to the complementary target RNA, followed byendonucleolytic cleavage. Specific ribozyme cleavage sites within apotential RNA target can be identified by known techniques. For ferdetails see, e.g., Rossi, Current Biologv, 4:469-471 (1994), and PCTpublication No. WO 97/33551 (published Sep. 18, 1997).

[1389] Nucleic acid molecules in triple-helix formation used to inhibittranscription should be single-stranded and composed ofdeoxynucleotides. The base composition of these oligonucleotides isdesigned such that it promotes triple-helix formation via Hoogsteenbase-pairing rules, which generally require sizeable stretches ofpurines or pyrimidines on one strand of a duplex. For farther detailssee, e.g., PCT publication No. WO 97/33551, supra.

[1390] These small molecules can be identified by any one or more of thescreening assays discussed hereinabove and/or by any other screeningtechniques well known for those skilled in the art.

[1391] PRO213 polypeptides and portions thereof which possess theability to regulate the growth induction cascade and/or the bloodcoagulation cascade may also be employed for such purposes both in vivotherapy and in vitro. Those of ordinary skill in the art will well knowhow to employ PRO213 polypeptides for such uses.

[1392] PRO274 polypeptides and portions thereof which have homology to7TM protein and Fn54 may also be useful for in vivo therapeuticpurposes, as well as for various other applications. The identificationof novel 7TM protein and Fn54like molecules may have relevance to anumber of human disorders which involve recognition of ligands and thesubsequent signal transduction of information contained within thoseligands in order to control cellular processes. Thus, the identificationof new 7TM protein and Fn54-like molecules is of special importance inthat such proteins may serve as potential therapeutics for a variety ofdifferent human disorders. Such polypeptides may also play importantroles in biotechnological and medical research as well as in variousindustrial applications. As a result, there is particular scientific andmedical interest in new molecules, such as PRO274.

[1393] PRO300 polypeptides and portions thereof which have homology toDiff 33 may also be useful for in vivo therapeutic purposes, as well asfor various other applications. The identification of novel Diff 33-likemolecules may have relevance to a number of human disorders such as thephysiology of cancer. Thus, the identification of new Diff 33-likemolecules is of special importance in that such proteins may serve aspotential therapeutics for a variety of different human disorders. Suchpolypeptides may also play important roles in biotechnological andmedical research as well as various industrial applications. As aresult, there is particular scientific and medical interest in newmolecules, such as PRO300.

[1394] PRO296 polypeptides of the present invention which possessbiological activity related to that of the sarcoma-amplified SAS proteinmay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO296 polypeptides of the present invention for such purposes.

[1395] PRO329 polypeptides of the present invention which possessbiological activity related to that of immunoglobulin F. receptorprotein or subunit thereof may be employed both in vivo for therapeuticpurposes and in vitro. Those of ordinary skill in the art will well knowhow to employ the PRO329 polypeptides of the present invention for suchpurposes.

[1396] PRO362 polypeptides of the present invention which possessbiological activity related to that of the A33 antigen protein, HCARprotein or the NrCAM related cell adhesion molecule may be employed bothin vivo for therapeutic purposes and in vitro.

[1397] PRO363 polypeptides of the present invention which possessbiological activity related to that of the cell surface HCAR protein maybe employed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO363polypeptides of the present invention for such purposes. Specifically,extracellular domains derived from the PRO363 polypeptides may beemployed therapeutically in vivo for lessening the effects of viralinfection.

[1398] PRO868 polypeptides of the present invention which possessbiological activity related to that of the tumor necrosis factor proteinmay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO868 polypeptides of the present invention for such purposes.

[1399] PRO382 polypeptides of the present invention which possessbiological activity related to that of the serine protease proteins maybe employed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO382polypeptides of the present invention for such purposes.

[1400] PRO545 polypeptides and portions thereof which have homology tomeltrin may also be useful for in vivo therapeutic purposes, as well asfor various other applications. The identification of novel moleculesassociated with cellular adhesion may be relevant to a number of humandisorders. Given that the meltrin proteins may play an important role ina number of disease processes, the identification of new meltrinproteins and meltrin-like molecules is of special importance in thatsuch proteins may serve as potential therapeutics for a variety ofdifferent human disorders. Such polypeptides may also play importantroles in biotechnological and medical research, as well as variousindustrial applications. As a result, there is particular scientific andmedical interest in new molecules, such as PRO545.

[1401] PRO617 polypeptides of the present invention which possessbiological activity related to that of the CD24 protein may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO617 polypeptides ofthe present invention for such purposes.

[1402] PRO700 polypeptides and portions thereof which have homology toprotein disulfide isomerase may also be useful for in vivo therapeuticpurposes, as well as for various other applications. The identificationof novel protein disulfide isomerases and related molecules may berelevant to a number of human disorders. Given that formation ofdisulfide bonds and protein folding play important roles in a number ofbiological processes, the identification of new protein disulfideisomerases and protein disulfide isomerase-like molecules is of specialimportance in that such proteins may serve as potential therapeutics fora variety of different human disorders. Such polypeptides may also playimportant roles in biotechnological and medical research, as well asvarious industrial applications. As a result, there is particularscientific and medical interest in new molecules, such as PRO700.

[1403] PRO702 polypeptides of the present invention which possessbiological activity related to that of the conglutinin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO702polypeptides of the present invention for such purposes. PRO702polypeptides having conglutinin activity would be expected to be capableof inhibiting haemagglutinin activity by influenza viruses and/orfunction as immunoglobulin-independent defense molecules as a result ofa complement-mediated mechanism.

[1404] PRO703 polypeptides of the present invention which possessbiological activity related to that of the VLCAS protein may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO703 polypeptides ofthe present invention for such purposes.

[1405] PRO703 polypeptides and portions thereof which have homology toVLCAS may also be useful for in vivo therapeutic purposes, as well asfor various other applications. The identification of novel VLCASproteins and related molecules may be relevant to a number of humandisorders. Thus, the identification of new VLCAS proteins and VLCASprotein-like molecules is of special importance in that such proteinsmay serve as potential therapeutics for a variety of different humandisorders. Such polypeptides may also play important roles inbiotechnological and medical research as well as various industrialapplications. As a result, there is particular scientific and medicalinterest in new molecules, such as PRO703.

[1406] PRO705 polypeptides of the present invention which possessbiological activity related to that of the K-glypican protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO705polypeptides of the present invention for such purposes.

[1407] PRO708 polypeptides of the present invention which possessbiological activity related to that of the aryl sulfatase proteins maybe employed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO708polypeptides of the present invention for such purposes.

[1408] PRO320 polypeptides of the present invention which possessbiological activity related to that of the fibulin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO320polypeptides of the present invention for such purposes.

[1409] PRO320 polypeptides and portions thereof which have homology tofibulin may also be useful for in vivo therapeutic purposes, as well asfor various other applications. The identification of novel fibulinproteins and related molecules may be relevant to a number of humandisorders such as cancer or those involving connective tissue,attachment molecules and related mechanisms. Thus, the identification ofnew fibulin proteins and fibulin protein-like molecules is of specialimportance in that such proteins may serve as potential therapeutics fora variety of different human disorders. Such polypeptides may also playimportant roles in biotechnological and medical research as well asvarious industrial applications. As a result, there is particularscientific and medical interest in new molecules, such as PRO320.

[1410] PRO324 polypeptides of the present invention which possessbiological activity related to that of oxidoreductases may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO324 polypeptides ofthe present invention for such purposes.

[1411] PRO351 polypeptides of the present invention which possessbiological activity related to that of the prostasin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO351polypeptides of the present invention for such purposes.

[1412] PRO351 polypeptides and portions thereof which have homology toprostasin may also be useful for in vivo therapeutic purposes, as wellas for various other applications. The identification of novel prostasinproteins and related molecules may be relevant to a number of humandisorders. Thus, the identification of new prostasin proteins andprostasin-like molecules is of special importance in that such proteinsmay serve as potential therapeutics for a variety of different humandisorders. Such polypeptides may also play important roles inbiotechnological and medical research as well as various industrialapplications. As a result, there is particular scientific and medicalinterest in new molecules, such as PRO351.

[1413] PRO352 polypeptides of the present invention which possessbiological activity related to that of the butyrophilin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO352polypeptides of the present invention for such purposes.

[1414] PRO381 polypeptides of the present invention which possessbiological activity related to that of one or more of the FKPBimmunophilin proteins may be employed both in vivo for therapeuticpurposes and in vitro, for example for enhancing immunosuppressantactivity and/or for axonal regeneration. Those of ordinary skill in theart will well know how to employ the PRO381 polypeptides of the presentinvention for such purposes.

[1415] PRO386 polypeptides of the present invention which possessbiological activity related to that of the beta-2 subunit of a sodiumchannel expressed in mammalian cells may be employed both in vivo fortherapeutic purposes and in vitro. Those of ordinary skill in the artwill well know how to employ the PRO386 polypeptides of the presentinvention for such purposes.

[1416] PRO540 polypeptides of the present invention which possessbiological activity related to that of the LCAT protein may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO540 polypeptides ofthe present invention for such purposes.

[1417] PRO615 polypeptides of the present invention which possessbiological activity related to that of the synaptogyrin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO615polypeptides of the present invention for such purposes.

[1418] PRO615 polypeptides and portions thereof which have homology tosynaptogyrin may also be useful for in vivo therapeutic purposes, aswell as for various other applications. The identification of novelsynaptogyrin proteins and related molecules may be relevant to a numberof human disorders. Thus, the identification of new synaptogyrinproteins and synaptogyrin-like molecules is of special importance inthat such proteins may serve as potential therapeutics for a variety ofdifferent human disorders. Such polypeptides may also play importantroles in biotechnological and medical research as well as variousindustrial applications. As a result, there is particular scientifican(d medical interest in new molecules, such as PRO615.

[1419] PRO618 polypeptides of the present invention which possessbiological activity related to that of an enteropeptidase may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO618polypeptides of the present invention for such purposes.

[1420] PRO618 polypeptides and portions thereof which have homology toenteropeptidase may also be useful for in vivo therapeutic purposes, aswell as for various other applications. The identification of novelenteropeptidase proteins and related molecules may be relevant to anumber of human disorders. Thus, the identification of newenteropeptidase proteins and enteropeptidase-like molecules is ofspecial importance in that such proteins may serve as potentialtherapeutics for a variety of different human disorders. Suchpolypeptides may also play important roles in biotechnological andmedical research as well as various industrial applications. As aresult, there is particular scientific and medical interest in newmolecules, such as PRO618.

[1421] PRO719 polypeptides of the present invention which possessbiological activity related to that of the lipoprotein lipase H proteinmay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO719 polypeptides of the present invention for such purposes.

[1422] PRO724 polypeptides of the present invention which possessbiological activity related to that of the human LDL receptor proteinmay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO724 polypeptides of the present invention for such purposes.

[1423] PRO772 polypeptides of the present invention which possessbiological activity related to that of the human A4 protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO772polypeptides of the present invention for such purposes.

[1424] PRO852 polypeptides of the present invention which possessbiological activity related to that of certain protease protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO852polypeptides of the present invention for such purposes.

[1425] PRO853 polypeptides of the present invention which possessbiological activity related to that of the reductase protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO853polypeptides of the present invention for such purposes.

[1426] PRO853 polypeptides and portions thereof which have homology toreductase proteins may also be useful for in vivo therapeutic purposes,as well as for various other applications. Given that oxygen freeradicals and antioxidants appear to play important roles in a number ofdisease processes, the identification of new reductase proteins andreductase-like molecules is of special importance in that such proteinsmay serve as potential therapeutics for a variety of different humandisorders. Such polypeptides may also play important roles inbiotechnological and medical research as well as various industrialapplications. As a result, there is particular scientific and medicalinterest in new molecules, such as PRO853.

[1427] PRO860 polypeptides of the present invention which possessbiological activity related to that of the neurofascin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO860polypeptides of the present invention for such purposes.

[1428] PRO860 polypeptides and portions thereof which have homology toneurofascin may also be useful for in vivo therapeutic purposes, as wellas for various other applications. The identification of novelneurofascin proteins and related molecules may be relevant to a numberof human disorders which involve cellular adhesion. Thus, theidentification of new neurofascin proteins and neurofascin protein-likemolecules is of special importance in that such proteins may serve aspotential therapeutics for a variety of different human disorders. Suchpolypeptides may also play important roles in biotechnological andmedical research as well as various industrial applications. As aresult, there is particular scientific and medical interest in newmolecules, such as PRO860.

[1429] PRO846 polypeptides of the present invention which possessbiological activity related to that of the CMRF35 protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO846polypeptides of the present invention for such purposes.

[1430] PRO846 polypeptides and portions thereof which have homology tothe CMRF35 protein may also be useful for in vivo therapeutic purposes,as well as for various other applications. The identification of novelCMRF35 protein and related molecules may be relevant to a number ofhuman disorders. Thus, the identification of new CMRF35 protein andCMRF35 protein-like molecules is of special importance in that suchproteins may serve as potential therapeutics for a variety of differenthuman disorders. Such polypeptides may also play important roles inbiotechnological and medical research as well as various industrialapplications. As a result, there is particular scientific and medicalinterest in new molecules, such as PRO846.

[1431] PRO862 polypeptides of the present invention which possessbiological activity related to that of the lysozyme protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO862polypeptides of the present invention for such purposes.

[1432] PRO862 polypeptides and portions thereof which have homology tothe lysozyme protein may also be useful for in vivo therapeuticpurposes, as well as for various other applications. The identificationof novel lysozyme proteins and related molecules may be relevant to anumber of human disorders. Thus, the identification of new lysozymes andlysozyme-like molecules is of special importance in that such proteinsmay serve as potential therapeutics for a variety of different humandisorders. Such polypeptides may also play important roles inbiotechnological and medical research as well as various industrialapplications. As a result, there is particular scientific and medicalinterest in new molecules, such as PRO862.

[1433] PRO864 polypeptides of the present invention which possessbiological activity related to that of the Wnt-4 protein may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO864 polypeptides ofthe present invention for such purposes.

[1434] PRO864 polypeptides and portions thereof which have homology tothe Wnt-4 protein may also be useful for in vivo therapeutic purposes,as well as for various other applications. The identification of novelWnt-4 proteins and related molecules may be relevant to a number ofhuman disorders. Thus, the identification of new Wnt-4 protein and Wnt-4protein-like molecules is of special importance in that such proteinsmay serve as potential therapeutics for a variety of different humandisorders. Such polypeptides may also play important roles inbiotechnological and medical research as well as various industrialapplications. As a result, there is particular scientific and medicalinterest in new molecules, such as PRO864.

[1435] PRO792 polypeptides of the present invention which possessbiological activity related to that of the CD23 protein may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO792 polypeptides ofthe present invention for such purposes.

[1436] PRO866 polypeptides of the present invention which possessbiological activity related to that of mindin and/or spondin protein maybe employed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO866polypeptides of the present invention for such purposes.

[1437] PRO871 polypeptides of the present invention which possessbiological activity related to that of the cyclophilin protein familymay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO871 polypeptides of the present invention for such purposes.

[1438] PRO873 polypeptides of the present invention which possessbiological activity related to that of carboxylesterases may be employedboth in vivo for therapeutic purposes and in vitro. For example, they beused in conjunction with prodrugs to convert the prodrug to its activeform (see Danks et al., supra). They may be used to inhibit parasiteinfection (see van Pelt et al., supra). Methods for employ the PRO873polypeptides of the present invention for these, and other purposes willbe readily apparent to those of ordinary skill in the art.

[1439] PRO940 polypeptides of the present invention which possessbiological activity related to that of the CD33 protein and/or OBbinding protein-2 may be employed both in vivo for therapeutic purposesand in vitro. Those of ordinary skiM in the art will well know how toemploy the PRO940 polypeptides of the present invention for suchpurposes.

[1440] PRO941 polypeptides of the present invention which possessbiological activity related to that of a cadherin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO941polypeptides of the present invention for such purposes.

[1441] PRO944 polypeptides of the present invention which possessbiological activity related to that of the CPE-R protein may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskil in the art will well know how to employ the PRO944 polypeptides ofthe present invention for such purposes. PRO944 polypeptides of thepresent invention that function to bind to Clostridium perfringensenterotoxin (CPE) may find use for effectively treating infection by theCPE endotoxin.

[1442] PRO983 polypeptides of the present invention which possessbiological activity related to that of the vesicle-associated membraneprotein, VAP-33, may be employed both in vivo for therapeutic purposesand in vitro. Those of ordinary skill in the art will well know how toemploy the PRO983 polypeptides of the present invention for suchpurposes.

[1443] PRO1057 polypeptides of the present invention which possessbiological activity related to that of protease proteins may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art win well know how to employ the PRO1057 polypeptides ofthe present invention for such purposes.

[1444] PRO1071 polypeptides of the present invention which possessbiological activity related to that of the thrombospondin protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO1071polypeptides of the present invention for such purposes.

[1445] PRO1072 polypeptides of the present invention which possessbiological activity related to that of reductase proteins may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO1072polypeptides of the present invention for such purposes.

[1446] PRO1075 polypeptides of the present invention which possessbiological activity related to that of protein disulfide isomerase maybe employed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO1075polypeptides of the present invention for such purposes.

[1447] PRO181 polypeptides of the present invention which possessbiological activity related to that of the cornichon protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO181polypeptides of the present invention for such purposes.

[1448] PRO827 polypeptides of the present invention which possessbiological activity related to that of various integrin proteins may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO827polypeptides of the present invention for such purposes.

[1449] PRO1114 polypeptides of the present invention which possessbiological activity related to that of the cytokine receptor family ofproteins may be employed both in vivo for therapeutic purposes and invitro. Those of ordinary skill in the art will well know how to employthe PRO1114 polypeptides of the present invention for such purposes.

[1450] In addition to the above, the PRO1114 interferon receptorpolypeptides may be employed in applications, both in vivo and in vitro,where the ability to bind to an interferon ligand is desired. Suchapplications will be well within the skill level in the art.

[1451] PRO237 polypeptides of the present invention which possessbiological activity related to that of the carbonic anhydrase proteinmay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO237 polypeptides of the present invention for such purposes.

[1452] PRO541 polypeptides of the present invention which possessbiological activity related to that of a trypsin inhibitor protein maybe employed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO541polypeptides of the present invention for such purposes.

[1453] PRO273 polypeptides can be used in assays that other chemokineswould be used in to perform comparative assays. The results can be usedaccordingly.

[1454] PRO701 polypeptides of the present invention which possessbiological activity related to that of the neuroligin family may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO701polypeptides of the present invention for such purposes.

[1455] PRO701 can be used in assays with neurons and its activitythereon can be compared with that of neuroligins 1, 2 and 3. The resultscan be applied accordingly.

[1456] PRO704 polypeptides of the present invention which possessbiological activity related to that of vesicular integral membraneproteins may be employed both in vivo for therapeutic purposes and invitro. Those of ordinary skill in the art will well know how to employthe PRO704 polypeptides of the present invention for such purposes.

[1457] PRO704 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly. PRO704 can be tagged or measured for activity tomeasure endocytosis activity and thereby used to screen for agents whicheffect endocytosis.

[1458] PRO706 polypeptides of the present invention which possessbiological activity related to that of the endogenous prostatic acidphosphatase precursor may be employed both in vivo for therapeuticpurposes and in vitro. Those of ordinary skill in the art will well knowhow to employ the PRO706 polypeptides of the present invention for suchpurposes.

[1459] PRO706 can be used in assays with human prostatic acidphosphatase or human lysosomal acid phosphatase and its activity thereoncan be compared with that of human prostatic acid phosphatase or humanlysosomal acid phosphatase. The results can be applied accordingly.

[1460] PRO707 polypeptides of the present invention which possessbiological activity related to that of cadherins may be employed both invivo for therapeutic purposes and in vitro. Those of ordinary skill inthe art will well know how to employ the PRO707 polypeptides of thepresent invention for such purposes.

[1461] PRO707 can be used in assays to determine its activity inrelation to other cadherins, particularly cadherin FIB3. The results canbe applied accordingly.

[1462] PRO322 polypeptides of the present invention which possessbiological activity related to that of neuropsin may be employed both invivo for therapeutic purposes and in vitro. Those of ordinary skill inthe art will well know how to employ the PRO322 polypeptides of thepresent invention for such purposes.

[1463] PRO322 can be used in assays to determine its activity relativeto neuropsin, trypsinogen, serine protease and neurosin, and the resultsapplied accordingly.

[1464] PRO526 polypeptides of the present invention which possessbiological activity related to that of protein-protein binding proteinsmay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO526 polypeptides of the present invention for such purposes.

[1465] Assays can be performed with growth factors and other proteinswhich are known to form complexes to determine whether PRO526 bindsthereto and whether there is increased half-life due to such binding.The results can be used accordingly.

[1466] PRO531 polypeptides of the present invention which possessbiological activity related to that of the protocadherins may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO531polypeptides of the present invention for such purposes.

[1467] PRO531 can be used in assays against protocadherin 3 and otherprotocadherins, to determine their relative activities. The results canbe applied accordingly.

[1468] PRO534 polypeptides of the present invention which possessbiological activity related to that of the protein disulfide isomerasemay be employed both in vivo for therapeutic purposes and in vitro.Those of ordinary skill in the art will well know how to employ thePRO534 polypeptides of the present invention for such purposes.

[1469] PRO534 can be used in assays with protein disulfide isomerase todetermine the relative activities. The results can be appliedaccordingly.

[1470] PRO697 polypeptides of the present invention which possessbiological activity related to that of the sFRP family may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO697 polypeptides ofthe present invention for such purposes.

[1471] PRO697 can be used in assays with sFRPs and SARPs to determinethe relative activities. The results can be applied accordingly.

[1472] PRO731 polypeptides of the present invention which possessbiological activity related to that of any protocadherin may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO731 polypeptides ofthe present invention for such purposes.

[1473] PRO731 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1474] PRO768 polypeptides of the present invention which possessbiological activity related to that of integrins may be employed both invivo for therapeutic purposes and in vitro. Those of ordinary skill inthe art will well know how to employ the PRO768 polypeptides of thepresent invention for such purposes.

[1475] PRO768 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1476] PRO771 polypeptides of the present invention which possessbiological activity related to that of the testican protein may beemployed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO771polypeptides of the present invention for such purposes.

[1477] PRO771 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1478] PRO733 polypeptides of the present invention which possessbiological activity related to that of the proteins which bind theT1/ST2 receptor may be employed both in vivo for therapeutic purposesand in vitro. Those of ordinary skill in the art will well know how toemploy the PRO733 polypeptides of the present invention for suchpurposes.

[1479] PRO733 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1480] PRO162 polypeptides of the present invention which possessbiological activity related to that of the pancreatitis-associatedprotein may be employed both in vivo for therapeutic purposes and invitro. Those of ordinary skill in the art will well know how to employthe PRO162 polypeptides of the present invention for such purposes.

[1481] PRO162 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1482] PRO788 polypeptides of the present invention which possessbiological activity related to that of the anti-neoplastic urinaryprotein may be employed both in vivo for therapeutic purposes and invitro. Those of ordinary skill in the art will well know how to employthe PRO788 polypeptides of the present invention for such purposes.

[1483] PRO788 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1484] PRO1008 polypeptides of the present invention which possessbiological activity related to that of dkk-1 may be employed both invivo for therapeutic purposes and in vitro. Those of ordinary skill inthe art will well know how to employ the PRO1008 polypeptides of thepresent invention for such purposes.

[1485] PRO1008 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1486] PRO1012 polypeptides of the present invention which possessbiological activity related to that of the protein disulfide isomerasemay be employed both in vivo and in vitro purposes. Those of ordinaryskill in the art will well know how to employ the PRO1012 polypeptidesof the present invention for such purposes.

[1487] PRO1012 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1488] PRO1014 polypeptides of the present invention which possessbiological activity related to that of reductase may be employed both invivo for therapeutic purposes and in vitro. Those of ordinary skill inthe art will well know how to employ the PRO1014 polypeptides of thepresent invention for such purposes.

[1489] PRO1014 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. Inhibitors ofPRO1014 are particularly preferred. The results can be appliedaccordingly.

[1490] PRO1017 polypeptides of the present invention which possessbiological activity related to that of sulfotransferase may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO1017 polypeptidesof the present invention for such purposes.

[1491] PRO1017 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1492] PRO474 polypeptides of the present invention which possessbiological activity related to that of dehydrogenase may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskid in the art will well know how to employ the PRO474 polypeptides ofthe present invention for such purposes.

[1493] PRO474 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1494] PRO1031 polypeptides of the present invention which possessbiological activity related to that of IL-17 may be employed both invivo for therapeutic purposes and in vitro. Those of ordinary skill inthe art will well know how to employ the PRO1031 polypeptides of thepresent invention for such purposes.

[1495] PRO1031 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly.

[1496] PRO938 polypeptides of the present invention which possessbiological activity related to that of protein disulfide isomerase maybe employed both in vivo for therapeutic purposes and in vitro. Those ofordinary skill in the art will well know how to employ the PRO938polypeptides of the present invention for such purposes.

[1497] PRO1082 polypeptides of the present invention which possessbiological activity related to that of the LDL receptor may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO1082 polypeptidesof the present invention for such purposes.

[1498] PRO1082 can be used in assays with the polypeptides to which theyhave identity with to determine the relative activities. The results canbe applied accordingly. PRO1082 can also be used in assays to identifycandidate agents which modulate the receptors.

[1499] PRO1083 polypeptides of the present invention which possessbiological activity related to that of 7TM receptors may be employedboth in vivo for therapeutic purposes and in vitro. Those of ordinaryskill in the art will well know how to employ the PRO1083 polypeptidesof the present invention for such purposes.

[1500] In particular PRO1083 can be used in assays to determinecandidate agents which control or modulate PRO1083, i.e., have an effecton the receptor.

[1501] The VEGF-E molecules herein have a number of therapeutic usesassociated with survival, proliferation and/or differention of cells.Such uses include the treatment of umbilical vein endothelial cells, inview of the demonstrated ability of VEGF-E to increase survival of humanumbilical vein endothelial cells. Treatment may be needed if the veinwere subjected to traumata, or situations wherein artificial means areemployed to enhance the survival of the umbilical vein, for example,where it is weak, diseased, based on an artificial matrix, or in anartificial environment. Other physiological conditions that could beimproved based on the selective mitogenic character of VEGF-E are alsoincluded herein. Uses also include the treatment of fibroblasts andmyocytes, in view of the demonstrated ability of VEGF-E to induceproliferation of fibroblasts and hypertrophy in myocytes. In particular,VEGF-E can be used in wound healing, tissue growth and muscle generationand regeneration.

[1502] For the indications referred to above, the VEGF-E molecule willbe formulated and dosed in a fashion consistent with good medicalpractice taking into account the specific disorder to be treated, thecondition of the individual patient, the site of delivery of the VEGF-E,the method of administration, and other factors known to practitioners.Thus, for purposes herein, the “therapeutically effective amount” of theVEGF-E is an amount that is effective either to prevent, lessen theworsening of, alleviate, or cure the treated condition, in particularthat amount which is sufficient to enhance the survival, proliferationand/or differentiation of the treated cells in vivo.

[1503] VEGF-E amino acid variant sequences and derivatives that areimmunologically crossreactive with antibodies raised against native VEGFare useful in immunoassays for VEGF-E as standards, or, when labeled, ascompetitive reagents.

[1504] The VEGF-E is prepared for storage or administration by mixingVEGF-E having the desired degree of purity with physiologicallyacceptable carriers, excipients, or stabilizers. Such materials arenon-toxic to recipients at the dosages and concentrations employed. Ifthe VEGF-E is water soluble, it may be formulated in a buffer such asphosphate or other organic acid salt preferably at a pH of about 7 to 8.If the VEGF-E is only partially soluble in water, it may be prepared asa microemulsion by formulating it with a nonionic surfactant such asTween, Pluronics, or PEG, e.g., Tween 80, in an amount of 0.04-0.05%(w/v), to increase its solubility.

[1505] Optionally other ingredients may be added such as antioxidants,e.g., ascorbic acid; low molecular weight (less than about ten residues)polypeptides, e.g., polyarginine or tripeptides; proteins, such as serumalbumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids, such as glycine, glutamic acid,aspartic acid, or arginine; monosaccharides, disaccharides, and othercarbohydrates including cellulose or its derivatives, glucose, marmose,or dextrins; chelating agents such as EDTA; and sugar alcohols such asmannitol or sorbitol.

[1506] The VEGF-E to be used for therapeutic administration must besterile. Sterility is readily accomplished by filtration through sterilefiltration membranes (e.g., 0.2 micron membranes). The VEGF-E ordinarilywin be stored in lyophilized form or as an aqueous solution if it ishighly stable to thermal and oxidative denaturation. The pH of theVEGF-E preparations typically will be about from 6 to 8, although higheror lower pH values may also be appropriate in certain instances. It willbe understood that use of certain of the foregoing excipients, carriers,or stabilizers will result in the formation of salts of the VEGF-E.

[1507] If the VEGF-E is to be used parenterally, therapeuticcompositions containing the VEGF-E generally are placed into a containerhaving a sterile access port, for example, an intravenous solution bagor vial having a stopper pierceable by a hypodermic injection needle.

[1508] Generally, where the disorder permits, one should formulate anddose the VEGF-E for site-specific delivery. This is convenient in thecase of wounds and ulcers. Sustained release formulations may also beprepared, and include the formation of microcapsular particles andimplantable articles. For preparing sustained-release VEGF-Ecompositions, the VEGF-E is preferably incorporated into a biodegradablematrix or microcapsule. A suitable material for this purpose is apolylactide, although other polymers of poly-(a-hydroxycarboxylicacids), such as poly-D-(−)-3-hydroxybutyric acid (EP 133,988A), can beused. Other biodegradable polymers include poly(lactones),poly(acetals), poly(orthoesters), or poly(orthocarbonates). The initialconsideration here must be that the carrier itself, or its degradationproducts, is nontoxic in the target tissue and will not furtheraggravate the condition. This can be determined by routine screening inanimal models of the target disorder or, if such models are unavailable,in normal animals. Numerous scientific publications document such animalmodels.

[1509] For examples of sustained release compositions, see U.S. Pat. No.3,773,919, EP 58,481A, U.S. Pat. No. 3,887,699, EP 158,277A, CanadianPatent No. 1176565, U. Sidman et al., Biopolymers 22, 547 [1983], and R.Langer et al., Chem. Tech. 12, 98 [1982].

[1510] When applied topically, the VEGF-E is suitably combined withother ingredients, such as carriers and/or adjuvants. There are nolimitations on the nature of such other ingredients, except that theymust be pharmaceutically acceptable and efficacious for their intendedadministration, and cannot degrade the activity of the activeingredients of the composition. Examples of suitable vehicles includeointments, creams, gels, or suspensions, with or without purifiedcollagen. The compositions also may be impregnated into transdermalpatches, plasters, and bandages, preferably in liquid or semi-liquidform.

[1511] For obtaining a gel formulation, the VEGF-E formulated in aliquid composition may be mixed with an effective amount of awater-soluble polysaccharide or synthetic polymer such as polyethyleneglycol to form a gel of the proper viscosity to be applied topically.The polysaccharide that may be used includes, for example, cellulosederivatives such as etherified cellulose derivatives, including alkylcelluloses, hydroxyalkyl celluloses, and alkylhydroxyalkyl celluloses,for example, methylcellulose, hydroxyethyl cellulose, carboxymethylcellulose, hydroxypropyl methylcellulose, and hydroxypropyl cellulose;starch and fractionated starch; agar; alginic acid and alginates;gumarabic; pullullan; agarose; carrageenan; dextrans; dextrans;fructans; inulin; mannans; xylans; arabinans; chitosans; glycogens;glucans; and synthetic biopolymers; as well as gums such as xanthan gum;guar gum; locust bean gum; gum arabic; tragacanth gum; and karaya gum;and derivatives and mixtures thereof. The preferred gelling agent hereinis one that is inert to biological systems, nontoxic, simple to prepare,and not too runny or viscous, and will not destabilize the VEGF-E heldwithin it.

[1512] Preferably the polysaccharide is an etherified cellulosederivative, more preferably one that is well defined, purified, andlisted in USP, e.g., methylcellulose and the hydroxyalkyl cellulosederivatives, such as hydroxypropyl cellulose, hydroxyethyl cellulose,and hydroxypropyl methylcellulose. Most preferred herein ismethylcellulose.

[1513] The polyethylene glycol useful for gelling is typically a mixtureof low and high molecular weight polyethylene glycols to obtain theproper viscosity. For example, a mixture of a polyethylene glycol ofmolecular weight 400-600 with one of molecular weight 1500 would beeffective for this purpose when mixed in the proper ratio to obtain apaste.

[1514] The term “water soluble” as applied to the polysaccharides andpolyethylene glycols is meant to include colloidal solutions anddispersions. In general, the solubility of the cellulose derivatives isdetermined by the degree of substitution of ether groups, and thestabilizing derivatives useful herein should have a sufficient quantityof such ether groups per anhydroglucose unit in the cellulose chain torender the derivatives water soluble. A degree of ether substitution ofat least 0.35 ether groups per anhydroglucose unit is generallysufficient. Additionally, the cellulose derivatives may be in the formof alkali metal salts, for example, the Li, Na, K, or Cs salts.

[1515] If methylcellulose is employed in the gel, preferably itcomprises about 2-5%, more preferably about 3%, of the gel and the VEGFis present in an amount of about 300-1000 mg per ml of gel.

[1516] The dosage to be employed is dependent upon the factors describedabove. As a general proposition, the VEGF-E is formulated and deliveredto the target site or tissue at a dosage capable of establishing in thetissue a VEGF-E level greater than about 0.1 ng/cc up to a maximum dosethat is efficacious but not unduly toxic. This intra-tissueconcentration should be maintained if possible by continuous infusion,sustained release, topical application, or injection at empiricallydetermined frequencies.

[1517] It is within the scope hereof to combine the VEGF-E therapy withother novel or conventional therapies (e.g., growth factors such asVEGF, aFGF, bFGF, PDGF, IGF, NGF, anabolic steroids, EGF or TGF-a) forenhancing the activity of any of the growth factors, including VEGF-E,in promoting cell proliferation, survival, differentiation and repair.It is not necessary that such cotreatment drugs be included per se inthe compositions of this invention, although this will be convenientwhere such drugs are proteinaceous. Such admixtures are suitablyadministered in the same manner and for the same purposes as the VEGF-Eused alone. The useful molar ratio of VEGF-E to such secondary growthfactors is typically 1:0.1-10, with about equimolar amounts beingpreferred.

[1518] The compounds of the present invention can be formulatedaccording to known methods to prepare pharmaceutically usefulcompositions, whereby the PRO polypeptide hereof is combined inadmixture with a pharmaceutically acceptable carrier vehicle. Suitablecarrier vehicles and their formulation, inclusive of other humanproteins, e.g., human serum albumin, are described, for example, inRemington's Pharmaceutical Sciences, 16th ed., 1980, Mack PublishingCo., edited by Oslo et aL the disclosure of which is hereby incorporatedby reference. The VEGF-E herein may be administered parenterally tosubjects suffering from cardiovascular diseases or conditions, or byother methods that ensure its delivery to the bloodstream in aneffective form.

[1519] Compositions particularly well suited for the clinicaladministration of VEGF-E hereof employed in the practice of the presentinvention include, for example, sterile aqueous solutions, or sterilehydratable powders such as lyophiaed protein. It is generally desirableto include further in the formulation an appropriate amount of apharmaceutically acceptable salt, generally in an amount sufficient torender the formulation isotonic. A pH regulator such as arginine base,and phosphoric acid, are also typically included in sufficientquantities to maintain an appropriate pH, generally from 5.5 to 7.5.Moreover, for improvement of shelf-life or stability of aqueousformulations, it may also be desirable to include further agents such asglycerol. hi this manner, variant t-PA formulations are renderedappropriate for parenteral administration, and, in particular,intravenous administration.

[1520] Dosages and desired drug concentrations of pharmaceuticalcompositions of the present invention may vary depending on theparticular use envisioned. For example, in the treatment of deep veinthrombosis or peripheral vascular disease, “bolus” doses, will typicallybe preferred with subsequent administrations being given to maintain anapproximately constant blood level, preferably on the order of about 3μg/ml.

[1521] However, for use in connection with emergency medical carefacilities where infusion capability is generally not available and dueto the generally critical nature of the underlying disease (e.g.,embolism, infarct), it will generally be desirable to provide somewhatlarger initial doses, such as an intravenous bolus.

[1522] For the various therapeutic indications referred to for thecompounds hereof, the VEGF-E molecules will be formulated and dosed in afashion consistent with good medical practice taking into account thespecific disorder to be treated, the condition of the individualpatient, the site of delivery, the method of administration and otherfactors known to practitioners in the respective art. Thus, for purposesherein, the “therapeutically effective amount” of the VEGF-E moleculeshereof is an amount that is effective either to prevent, lessen theworsening of, alleviate, or cure the treated condition, in particularthat amount which is sufficient to enhance the survival, proliferationor differentiation of targeted cells in vivo. In general a dosage isemployed capable of establishing in the tissue that is the target forthe therapeutic indication being treated a level of a VEGF-E hereofgreater than about 0.1 ngcm³ up to a maximum dose that is efficaciousbut not unduly toxic. It is contemplated that intra-tissueadministration may be the choice for certain of the therapeuticindications for the compounds hereof.

[1523] The human Toll proteins of the present invention can also be usedin assays to identify other proteins or molecules involved inToll-mediated signal transduction. For example, PRO285 and PRO286 areuseful in identifying the as of yet unknown natural ligands of humanTolls, or other factors that participate (directly or indirectly) in theactivation of and/or signaling through a human Toll receptor, such aspotential Toll receptor associated kinases. In addition, inhibitors ofthe receptor/ligand binding interaction can be identified. Proteinsinvolved in such binding interactions can also be used to screen forpeptide or small molecule inhibitors or agonists of the bindinginteraction. Screening assays can be designed to find lead compoundsthat mimic the biological activity of a native Toll polypeptide or aligand for a native Toll polypeptide. Such screening assays will includeassays amenable to high-throughput screening of chemical libraries,making them particularly suitable for identifying small molecule drugcandidates. Small molecules contemplated include synthetic organic orinorganic compounds. The assays can be performed in a variety offormats, including protein-protein binding assays, biochemical screeningassays, immunoassays and cell based assays, which are well characterizedin the art.

[1524] In vitro assays employ a mixture of components including a Tollreceptor polypeptide, which may be part of fusion product with anotherpeptide or polypeptide, e.g., a tag for detecting or anchoring, etc. Theassay mixtures may further comprise (for binding assays) a naturalintra- or extracellular Toll binding target (i.e. a Toll ligand, oranother molecule known to activate and/or signal through the Tollreceptor). While native binding targets may be used, it is frequentlypreferred to use portion of such native binding targets (e.g. peptides),so long as the portion provides binding affinity and avidity to thesubject Toll protein conveniently measurable in the assay. The assaymixture also contains a candidate pharmacological agent. Candidateagents encompass numerous chemical classes, through typically they areorganic compounds, preferably small organic compounds, and are obtainedfrom a wide variety of sources, including libraries of synthetic ornatural compounds. A variety of other reagents may also be included inthe mixture, such as, salts, buffers, neutral proteins, e.g. albumin,detergents, protease inhibitors, nuclease inhibitors, antimicrobialagents, etc.

[1525] In in vitro binding assays, the resultant mixture is incubatedunder conditions whereby, but for the presence of the candidatemolecule, the Toll protein specifically binds the cellular bindingtarget, portion or analog, with a reference binding affinity. Themixture components can be added in any order that provides for therequisite bindings and incubations may be performed at any temperaturewhich facilitates optimal binding. Incubation periods are likewiseselected for optimal binding but also miniized to facilitate rapidhigh-throughput screening.

[1526] After incubation, the agent-biased binding between the Tollprotein and one or more binding targets is detected by any convenienttechnique. For cell-free binding type assays, a separation step is oftenused to separate bound from unbound components. Separation may beeffected by precipitation (e.g. TCA precipitation, immunoprecipitation,etc.), immobilization (e.g. on a solid substrate), etc., followed bywashing by, for example, membrane filtration (e.g. Whatman's P-18 ionexchange paper, Polyfiltronic's hydrophobic GFC membrane, etc.), gelchromatography (e.g. gel filtration, affinity, etc.). For Toll-dependenttranscription assays, binding is detected by a change in the expressionof a Toll-dependent reporter.

[1527] Detection may be effected in any convenient way. For cell-freebinding assays, one of the components usually comprises or is coupled toa label. The label may provide for direct detection as radioactivity,luminescence, optical or electron density, etc., or indirect detection,such as, an epitope tag, an enzyme, etc. A variety of methods may beused to detect the label depending on the nature of the label and otherassay components, e.g. through optical or electron density, radiativeemissions, nonradiative energy transfers, etc. or indirectly detectedwith antibody conjugates, etc.

[1528] Nucleic acid encoding the Toll polypeptides disclosed herein mayalso be used in gene therapy. In gene therapy applications, genes areintroduced into cells in order to achieve in vivo synthesis of atherapeutically effective genetic product, for example for replacementof a defective gene. “Gene therapy” includes both -conventional genetherapy where a lasting effect is achieved by a single treatment, andthe administration of gene H therapeutic agents, which involves the onetime or repeated administration of a therapeutically effective DNA ormRNA. Antisense RNAs and DNAs can be used as therapeutic agents forblocking the expression of certain genes in vivo. It has already beenshown that short antisense oligonucleotides can be imported into cellswhere they act as inhibitors, despite their low intracellularconcentrations caused by their restricted uptake by the cell membrane.(Zamecnik et al., Proc. Natl. Acad. Sci. USA 83, 4143-4146 [1986]). Theoligonucleotides can be modified to enhance their uptake, e.g. bysubstituting their negatively charged phosphodiester groups by unchargedgroups.

[1529] There are a variety of techniques available for introducingnucleic acids into viable cells. The techniques vary depending uponwhether the nucleic acid is transferred into cultured cells in vitro, orin vivo in the cells of the intended host. Techniques suitable for thetransfer of nucleic acid into mammalian cells in vitro include the useof liposomes, electroporation, microinjection, cell fusion,DEAE-dextran, the calcium phosphate precipitation method, etc. Thecurrently preferred in vivo gene transfer techniques includetransfection with viral (typically retroviral) vectors and viral coatprotein-liposome mediated transfection (Dzau et al., Trends inBiotechnology 11, 205-210 [1993]). In some situations it is desirable toprovide the nucleic acid source with an agent that targets the targetcells, such as an antibody specific for a cell surfae, membrane proteinor the target cell, a ligand for a receptor on the target cell, etc.Where liposomes are employed, proteins which bind to a cell surfacemembrane protein associated with endocytosis may be used for targetingand/or to facilitate uptake, e.g. capsid proteins or fragments thereoftropic for a particular cell type, antibodies for proteins which undergointernalization in cycling, proteins that target intracellularlocalization and enhance intracellular half-life. The technique ofreceptor-mediated endocytosis is described, for example, by Wu et al.,J. Biol. Chem. 262, 4429-4432 (1987); and Wagner et al., Proc. Natl.Acad. Sci. USA 87, 3410-3414 (1990). For review of the currently knowngene marking and gene therapy protocols see Anderson et al., Science256, 808-813 (1992).

[1530] The various uses listed in connection with the Toll proteinsherein, are also available for agonists of the native Toll receptors,which mimic at least one biological function of a native Toll receptor.

[1531] Neurotrimin as well as other members of the IgLON subfamily ofthe immunoglobulin superfamily have been identified to have effect uponneural patterning, differentiation, maturation and growth. As a result,PRO337 the human neurotrmin homolog polypeptides would be expected tohave utility in diseases which are characterized by neural disfunction.For example, motoneuron disorders such as amyotrophic lateral sclerosis(Lou Gehrig's disease), Bell's palsy, and various conditions involvingspinal muscular atrophy, or paralysis. NGF variant formulations of theinvention can be used to treat human neurodegenerative disorders, suchas Alzheimer's disease, Parkinson's disease, epilepsy, multiplesclerosis, Huntington's chorea, Down's Syndrome, nerve deafness, andMeniere's disease. Moreover PRO337 polypeptide may also be used as acognitive enhancer, to enhance learning particularly in dementia ortrauma, such as those associated with the above diseases.

[1532] Further, PRO337 may be employed to treat neuropathy, andespecially peripheral neuropathy. “Peripheral neuropathy” refers to adisorder affecting the peripheral nervous system, most often manifestedas one or a combination of motor, sensory, sensorimotor, or autonomicneural dysfunction. The wide variety of morphologies exhibited byperipheral neuropathies can each be attributed uniquely to an equallywide number of causes. For example, peripheral neuropathies can begenetically acquired, can result from a systemic disease, or can beinduced by a toxic agent. Examples include but are not limited todiabetic peripheral neuropathy, distal sensorimotor neuropathy, orautonomic neuropathies such as reduced motility of the gastrointestinaltract or atony of the urinary bladder. Examples of neuropathiesassociated with systemic disease include post-polio syndrome orAIDS-associated neuropathy; examples of hereditary neuropathies includeCharcot-Marie-Tooth disease, Refsum's disease, Abetalipoproteinemia,Tangier disease, Krabbe's disease, Metachromatic leukodystrophy, Fabry'sdisease, and Dejerine-Sottas syndrome; and examples of neuropathiescaused by a toxic agent include those caused by treatment with achemotherapeutic agent such as vincristine, cisplatin, methotrexate, or3′-azido-3′-deoxythymidine. Correspondingly, neurotrimin antagonistswould be expected to have utility in diseases characterized by excessiveneuronal activity.

[1533] Endothelin is generated from inactive intermediates, the bigendothelins, by a unique processing event catalyzed by the zincmetalloprotease, endothelin converting enzyme (ECE). ECE was recentlycloned, and its structure was shown to be a single pass transmembraneprotein with a short intracellular N-terminal and a long extracellularC-terminal that contains the catalytic domain and numerousN-glycosylation sites. ECEs cleave the endothelin propeptide betweenTrp73 and Val74 producing the active peptide, ET, which appears tofunction as a local rather than a circulating hormone (Rubanyi, G. M. &Polokoff, M. A., Pharmachological Reviews 46: 325-415 (1994). Thus ECEactivity is a potential site of regulation of endothelin production anda possible target for therapeutic intervention in the endothelin system.By blocking ECE activity, it is possible stop the production of ET-1 byinhibiting the conversion of the relatively inactive precursor, bigET-1, to the physiologically active form.

[1534] ECE-2 is 64% identical to bovine ECE-2 at the amino acid level.ECE-2 is closely related to ECE-1 (63% identical, 80% conserved),neutral endopeptidase 24.11 and the Kell blood group protein. BovineECE-2 is a type II membrane-bound metalloproteinase localized in thetrans-Golgi network where it acts as an intracellular enzyme convertingendogenous big endothelin-1 into active endothelin (Emoto, N. andYanangisawa, M., J. Biol. Chem. 270: 15262-15268 (1995). The bovineECE-2 mRNA expression is highest in parts of the brain, cerebral cortex,cerebellum and adrenal medulla. It is expressed at lower levels inmymetrium, testes, ovary, and endothelial cells. Bovine ECE-2 and ECE-1both are more active on ET-1 as a substrate compared to ET-2 or ET-3,Emoto and Yanangisawa, supra. Human ECE-2 is 736 amino acids in lengthwith a 31 residue amino-terminal tail, a 23 residue transmembrane helixand a 682 carboxy-terminal domain. It is 94% identical to bovine ECE-2and 64% identical to human ECE-1. The predicted transmembrane domain ishighly conserved between the human and bovine ECE-2 proteins and betweenhuman ECE-1 and human ECE-2, as are the putative N-linked glycosylationsites, Cys residues conserved in the neutral endopeptidase 24.11 and theKell blood group protein family and the putative zinc binding motif. Thesequence suggests, that like other members of the NEP-ECE-Kell family,human ECE-2 encodes a type II transmembrane zinc-bindingmetalloproteinase, which, by extrapolation from what is known aboutbovine ECE-2, is an intracellular enzyme located within the secretorypathway which processes endogenously produced big ET-1 while it is stillin the secretory vesicles. Emoto and Yanangisawa, supra.

[1535] The expression pattern of ECE-2 differs from that observed forECE-1. Northern blot analysis of mRNA levels indicated low levels ofexpression of a 3.3 kb transcript in adult brain (highest in thecerebellum, putamen, medulla and temporal lobe, and lower in thecerebral cortex, occipital lobe and frontal lobe), spinal cord, lung andpancreas and higher levels of a 4.5 kb transcript in fetal brain andkidney. The two transcript sizes probably represent the use ofalternative polyadenylation sites as has been observed for bovine ECE-2(Emoto and Yanangisawa, supra) and ECE-1 (Xu et al., Cell 78: 473-485(1994). PCR on cDNA libraries indicated low levels of expression infetal brain, fetal kidney, fetal small intestine and adult testis. Fetalliver, fetal lung and adult pancreas were all negative.

[1536] The endothelin (ET) family of peptides have potent vascular,cardiac and renal actions which may be of pathophysiological importancein many human disease states. ET-1 is expressed as an inactive 212 aminoacid prepropeptide. The prepropeptide is first cleaved at Arg52-CysS3and Arg92-Ala93 and then the carboxy terminal Lys9l and Arg92 aretrimmed from the protein to generate the propeptide big ET-1. ECEs thencleave the propeptide between Trp73 and Val74, producing the activepeptide, ET, which appears to function as a local rather than acirculating hormone (Rubanyi and Polokoff, Pharma. R. 46: 325-415(1994).

[1537] Endothelins may play roles in the pathophysiology of a number ofdisease states including: 1) cardiovascular diseases (vasospasm,hypertension, myocardial ischemia; reperfusion injury and acutemyochardial infarction, stroke (cerebral ischemia), congestive heartfailure, shock, atherosclerosis, vascular thickening); 2) kidney disease(acute and chronic renal failure, glomerulonephritis, cirrhosis); 3)lung disease (bronchial asthma, pulmonary hypertension); 4)gastrointestinal disorders (gastric ulcer, inflammatory bowel diseases);5) reproductive disorders (premature labor, dysmenorhea, preeclampsia)and 6) carcinogenesis. Rubanyi & Polokoff, supra.

[1538] Diseases can be evaluated for the impact of ET upon them byexamining: 1) increased production of ETs; 2) increased reactivity toETs; and/or 3) efficacy of an ET receptor antagonist, antibody or ECEinhibitor. Response to the previous criteria suggest that ETs likelyplay roles in cerebral vasospasm following subarachnoid hemorrhage,hypertension (fulminant complications), acute renal failure andcongestive heart failure. While inhibitors of ET production or activityhave not been used in models of coronary vasospasm, acute myocardialinfarction, and atherosclerosis, they do have elevated ET levels andincrease reactivity to ETs. Shock and pulmonary hypertension alsoexhibit elevated ET levels (Rubanyi and Polokoff, supra). Inhibition ofECEs in these conditions may be of therapeutic value.

[1539] The expression pattern of ECE-2 differs from that observed forECE-1. ECE-2 was observed at low levels in the adult brain, lung andpancreas and higher levels in fetal brain and kidney by Northern blotanalysis (FIG. 8). PCR revealed low levels of expression in additionaltissues: fetal lung, fetal small intestine and adult testis. Fetal liverwas negative. A similar pattern was reported for bovine ECE-2 (Emoto andYanangisawa, supra). It is expressed in brain tissues (cerebral cortex,cerebellum and adrenal medulla), myometrium and testis, and in lowlevels in ovary and very low levels in many other tissues. Bovine ECE-1(Xu et al, supra) is more widely and more abundantly expressed. It isobserved in vascular endothelial cells of most organs and in someparenchymal cells. With the exception for brain, bovine ECE-2 mRNA waspresent at lower levels than ECE-1. Applicants believe ECE-2 to be aparticularly good target for the therapeutic intervention for diseasessuch as cerebral vasospasm following subarachnoid hemorrhage and stroke.

[1540] Uses of the herein disclosed molecules may also be based upon thepositive functional assay hits disclosed and described below.

[1541] F. Anti-PRO Antibodies

[1542] The present invention further provides anti-PRO antibodies.Exemplary antibodies include polyclonal, monoclonal, humanized,bispecific, and heteroconjugate antibodies.

[1543] 1. Polyclonal Antibodies

[1544] The anti-PRO antibodies may comprise polyclonal antibodies.Methods of preparing polyclonal antibodies are known to the skilledartisan. Polyclonal antibodies can be raised in a mammal, for example,by one or more injections of an immunizing agent and, if desired, anadjuvant. Typically, the immunizing agent and/or adjuvant will beinjected in the mammal by multiple subcutaneous or intraperitonealinjections. The immunizing agent may include the PRO polypeptide or afusion protein thereof. It may be useful to conjugate the immunizingagent to a protein known to be immunogenic in the mammal beingimmunized. Examples of such immunogenic proteins include but are notlimited to keyhole limpet hemocyanin, serum albumin, bovinethyroglobulin, and soybean trypsin inhibitor. Examples of adjuvantswhich may be employed include Freund's complete adjuvant and MPL-TDMadjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).The immunization protocol may be selected by one skilled in the artwithout undue experimentation.

[1545] 2. Monoclonal Antibodies

[1546] The anti-PRO antibodies may, alternatively, be monoclonalantibodies. Monoclonal antibodies may be prepared using hybridomamethods, such as those described by Kohler and Milstein, Nature, 256:495(1975). In a hybridoma method, a mouse, hamster, or other appropriatehost animal, is typically immunized with an immunizing agent to elicitlymphocytes that produce or are capable of producing antibodies thatwill specifically bind to the immunizing agent. Alternatively, thelymphocytes may be immunized in vitro.

[1547] The immunizing agent will typically include the PRO polypeptideor a fusion protein thereof. Generally, either peripheral bloodlymphocytes (“PBLs”) are used if cells of human origin are desired, orspleen cells or lymph node cells are used if non-human mammalian sourcesare desired. The lymphocytes are then fused with an immortalized cellline using a suitable fusing agent, such as polyethylene glycol, to forma hybridoma cell [Goding, Monoclonal Antibodies: Principles andPractice, Academic Press, (1986) pp. 59-103]. Immortalized cell linesare usually transformed mammalian cells, particularly myeloma cells ofrodent, bovine and human origin. Usually, rat or mouse myeloma celllines are employed. The hybridoma cells may be cultured in a suitableculture medium that preferably contains one or more substances thatinhibit the growth or survival of the unfused, immortalized cells. Forexample, if the parental cells lack the enzyme hypoxanthine guaninephosphoribosyl transferase (HGPRT or HPRT), the culture medium for thehybridomas typically will include hypoxanthine, aminopterin, andthymidine (“HAT medium), which substances prevent the growth ofHGPRT-deficient cells.

[1548] Preferred immortalized cell lines are those that fuseefficiently, support stable high level expression of antibody by theselected antibody-producing cells, and are sensitive to a medium such asHAT medium. More preferred immortalized cell lines are murine myelomalines, which can be obtained, for instance, from the Salk Institute CellDistribution Center, San Diego, Calif. and the American Type CultureCollection, Manassas, Va. Human myeloma and mouse-human heteromyelomacell lines also have been described for the production ofhumanmonoclonalantibodies [Kozbor, J. Immunol., 133:3001 (1984); Brodeuretal.,Monoclonal Antibody Production Techniques and Applications, MarcelDekker, Inc., New York, (1987) pp. 51-631.

[1549] The culture medium in which the hybridoma cells are cultured canthen be assayed for the presence of monoclonal antibodies directedagainst PRO. Preferably, the binding specificity of monoclonalantibodies produced by the hybridoma cells is determined byinmunoprecipitation or by an in vitro binding assay, such asradioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).Such techniques and assays are known in the art. The binding affinity ofthe monoclonal antibody can, for exairple, be determined by theScatchard analysis of Munson and Pollard, Anal. Biochem., 107:220(1980).

[1550] After the desired hybridoma cells are identified, the clones maybe subcloned by limiting dilution procedures and grown by standardmethods [Goding, supra]. Suitable culture media for this purposeinclude, for example, Dulbecco's Modified Eagle's Medium andRPMI-1640medium. Alternatively, the hybridomacells may be grown in vivo asascites in a mammal.

[1551] The monoclonal antibodies secreted by the subclones may beisolated or purified from the culture medium or ascites fluid byconventional immunoglobulin purification procedures such as, forexample, protein A-Sepharose, hydroxylapatite chromatography, gelelectrophoresis, dialysis, or affinity chromatography.

[1552] The monoclonal antibodies may also be made by recombinant DNAmethods, such as those described in U.S. Pat. No. 4,816,567. DNAencoding the monoclonal antibodies of the invention can be readilyisolated and sequenced using conventional procedures (e.g., by usingoligonucleotide probes that are capable of binding specifically to genesencoding the heavy and light chains of murine antibodies). The hybridomacells of the invention serve as a preferred source of such DNA. Onceisolated, the DNA may be placed into expression vectors, which are thentransfected into host cells such as simian COS cells, Chinese hamsterovary (CHO) cells, or myeloma cells that do not otherwise produceimmunoglobulin protein, to obtain the synthesis of monoclonal antibodiesin the recombinant host cells. The DNA also may be modified, forexample, by substituting the coding sequence for human heavy and lightchain constant domains in place of the homologous murine sequences [U.S.Pat. No. 4,816,567; Morrison et al., supra] s or by covalently joiningto the immunoglobulin coding sequence all or part of the coding sequencefor a non-immunoglobulin polypeptide. Such a non-immunoglobulinpolypeptide can be substituted for the constant domains of an antibodyof the invention, or can be substituted for the variable domains of oneantigen-combining site of an antibody of the invention to create achimeric bivalent antibody.

[1553] The antibodies may be monovalent antibodies. Methods forpreparing monovalent antibodies are well known in the art. For example,one method involves recombinant expression of immunoglobulin light chainand modified heavy chain. The heavy chain is truncated generally at anypoint in the Fc region so as to prevent heavy chain crosslinking.Alternatively, the relevant cysteine residues are substituted withanother amino acid residue or are deleted so as to prevent crosslinking.

[1554] In vitro methods are also suitable for preparing monovalentantibodies. Digestion of antibodies to produce fragments thereof,particularly, Fab fragments, can be accomplished using routinetechniques known in the art.

[1555] 3. Human and Humanized Antibodies

[1556] The anti-PRO antibodies of the invention may further comprisehumanized antibodies or human antibodies. Humanized forms of non-human(e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulinchains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)₂ or otherantigen-binding subsequences of antibodies) which contain minimalsequence derived from non-human immunoglobulin. Humanized antibodiesinclude human immunoglobulins (recipient antibody) in which residuesfrom a complementary determining region (CDR) of the recipient arereplaced by residues from a CDR of a non-human species (donor antibody)such as mouse, rat or rabbit having the desired specificity, affinityand capacity. In some instances, Fv framework residues of the humanimmunoglobulin are replaced by corresponding non-human residues.Humanized antibodies may also comprise residues which are found neitherin the recipient antibody nor in the imported CDR or frameworksequences. In general, the humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the CDR regions correspond to thoseof a non-human immunoglobulin and all or substantially all of the FRregions are those of a human immunoglobulin consensus sequence. Thehumanized antibody optimally also will comprise at least a portion of animmunoglobulin constant region (Fc), typically that of a humanimmunoglobulin [Jones et al., Nature, 321:522-525 (1986); Riechmann etal., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol.,2:593-596 (1992)].

[1557] Methods for humanizing non-human antibodies are well known in theart. Generally, a humanized antibody has one or more amino acid residuesintroduced into it from a source which is non-human. These non-humanamino acid residues are often referred to as “import” residues, whichare typically taken from an “import” variable domain. Humanization canbe essentially performed following the method of Winter and co-workers[Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature,332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)], bysubstituting rodent CDRs or CDR sequences for the correspondingsequences of ahumanantibody. Accordingly, such “humanized” antibodiesare chimeric antibodies (U.S. Pat. No. 4,816,567), wherein substantiallyless than an intact human variable domain has been substituted by thecorresponding sequence from a non-human species. In practice, humanizedantibodies are typically human antibodies in which some CDR residues andpossibly some FR residues are substituted by residues from analogoussites in rodent antibodies.

[1558] Human antibodies can also be produced using various techniquesknown in the art, including phage display libraries [Hoogenboom andWinter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol.,222:581 (1991)]. The techniques of Cole et al. and Boerner et al. arealso available for the preparation of human monoclonal antibodies (Coleet al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77(1985) and Boerner et al., J. Immunol., 147(1):86-95 (1991)]. Similarly,human antibodies can be made by introducing of human immunoglobulin lociinto transgenic animals, e.g., mice in which the endogenousimmunoglobulin genes have been partially or completely inactivated. Uponchallenge, human antibody production is observed, which closelyresembles that seen in humans in all respects, including generearrangement, assembly, and antibody repertoire. This approach isdescribed, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806;5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the followingscientific publications: Marks et al., Bio/Technology 10, 779-783(1992); Lonberg et al., Nature 368 856-859 (1994); Morrison, Nature 368,812-13 (1994); Fishwild et al., Nature Biotechnology 14, 845-51 (1996);Neuberger, Nature Biotechnology 14, 826 (1996); Lonberg and Huszar,Intern. Rev. Immunol. 13 65-93 (1995).

[1559] 4. Bispecific Antibodies

[1560] Bispecific antibodies are monoclonal, preferably human orhumanized, antibodies that have binding specificities for at least twodifferent antigens. In the present case, one of the bindingspecificities is for the PRO, the other one is for any other antigen,and preferably for a cell-surface protein or receptor or receptorsubunit.

[1561] Methods for making bispecific antibodies are known in the art.Traditionally, the recombinant production of bispecific antibodies isbased on the co-expression of two immunoglobulin heavy-chain/light-chainpairs, where the two heavy chains have different specificities [Miisteinand Cuello, Nature, 305:537-539 (1983)]. Because of the randomassortment of immunoglobulin heavy and light chains, these hybridomas(quadromas) produce a potential mixire of ten different antibodymolecules, of which only one has the correct bispecific structure. Thepurification of the correct molecule is usually accomplished by affinitychromatography steps. Similar procedures are disclosed in WO 93/08829,published May 13, 1993, and in Traunecker et al., EMBO J. 10:3655-3659(1991).

[1562] Antibody variable domains with the desired binding specificities(antibody-antigen combining sites) can be fused to immunoglobulinconstant domain sequences. The fusion preferably is with animmunoglobulin heavy-chain constant domain, comprising at least part ofthe hinge, CH2, and CH3 regions. It is preferred to have the firstheavy-chain constant region (CH1) containing the site necessary forlight-chain binding present in at least one of the fusions. DNAsencoding the immunoglobulin heavy-chain fusions and, if desired, theimmunoglobulin light chain, are inserted into separate expressionvectors, and are co-transfected into a suitable host organism. Forfurther details of generating bispecific antibodies see, for example,Suresh et al., Methods in Enzymology, 121:210 (1986).

[1563] According to another approach described in WO 96127011, theinterface between a pair of antibody molecules can be engineered tomaximize the percentage of heterodimers which are recovered fromrecombinant cell culture. The preferred interface comprises at least apart of the CH3 region of an antibody constant domain. In this method,one or more small amino acid side chains from the interface of the firstantibody molecule are replaced with larger side chains (e.g. tyrosine ortryptophan). Compensatory “cavities” of identical or similar size to thelarge side chain(s) are created on the interface of the second antibodymolecule by replacing large amino acid side chains with smaller ones(e.g. alanine or threonine). This provides a mechanism for increasingthe yield of the heterodimer over other unwanted end-products such ashomodimers.

[1564] Bispecific antibodies can be prepared as full length antibodiesor antibody fragments (e.g. F(ab′)₂ bispecific antibodies). Techniquesfor generating bispecific antibodies from antibody fragments have beendescribed in the literature. For example, bispecific antibodies can beprepared can be prepared using chemical linkage. Brennan et al., Science229:81 (1985) describe aprocedure wherein intact antibodies areproteolytically cleaved to generate F(ab′)₂ fragments. These fragmentsare reduced in the presence of the dithiol complexing agent sodiumarsenite to stabilize vicinal dithiols and prevent intermoleculardisulfide formation. The Fab′ fragments generated are then converted tothionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives isthen reconverted to the Fab′-thiol by reduction with mercaptoethylamineand is mixed with an equimolar amount of the other Fab′-TNB derivativeto form the bispecific antibody. The bispecific antibodies produced canbe used as agents for the selective immobilization of enzymes.

[1565] Fab′ fragments may be directly recovered from E. coli andchemically coupled to form bispecific antibodies. Shalaby et al., J.Exp. Med. 175:217-225 (1992) describe the production of a fullyhumanized bispecific antibody F(ab′)₂ molecule. Each Fab′ fragment wasseparately secreted from E. coil and subjected to directed chemicalcoupling in vitro to form the bispecific antibody. The bispecificantibody thus formed was able to bind to cells overexpressing the ErbB2receptor and normal human T cells, as well as trigger the lytic activityof human cytotoxic lymphocytes against human breast tumor targets.

[1566] Various technique for making and isolating bispecific antibodyfragments directly from recombinant cell culture have also beendescribed. For example, bispecific antibodies have been produced usingleucine zippers. Kostelny et al., J. Immunol. 148(5):1547-1553 (1992).The leucine zipper peptides from the Fos and Jun proteins were linked tothe Fab′ portions of two different antibodies by gene fusion. Theantibody homodimers were reduced at the hinge region to form monomersand then re-oxidized to form the antibody heterodimers. This method canalso be utilized for the production of antibody homodimers. The'diabody” technology described by Hollinger et al., Proc. Natl. Acad.Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism formaking bispecific antibody fragments. The fragments comprise aheavy-chain variable domain (V_(H)) connected to a light-chain variabledomain (V_(L)) by a linker which is too short to allow pairing betweenthe two domains on the same chain. Accordingly, the V_(H) and V_(L)domains of one fragment are forced to pair with the complementary V_(L)and V_(H) domains of another fragment, thereby forming twoantigen-binding sites. Another strategy for making bispecific antibodyfragments by the use of single-chain Fv (sFv) dimers has also beenreported. See, Gruber et al., J. Immunol. 152:5368 (1994). Antibodieswith more tan two valencies are contemplated. For example, trispecificantibodies can be prepared. Tutt et al., J. Immunol. 147:60 (1991).

[1567] Exemplary bispecific antibodies may bind to two differentepitopes on a given PRO polypeptide herein. Alternatively, an anti-PROpolypeptide arm may be combined with an arm which binds to a triggeringmolecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2,CD3, CD28, or B7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64),FcγRII (CD32) and FcγRM (CD16) so as to focus cellular defensemechanisms to the cell expressing the particular PRO polypeptide.Bispecific antibodies may also be used to localize cytotoxic agents tocells which express a particular PRO polypeptide. These antibodiespossess a PRO-binding arm and an arm which binds a cytotoxic agent or aradionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Anotherbispecific antibody of interest binds the PRO polypeptide and furtherbinds tissue factor (TF).

[1568] 5. Heteroconjugate Antibodies

[1569] Heteroconjugate antibodies are also within the scope of thepresent invention. Heteroconjugate antibodies are composed of twocovalenily joined antibodies. Such antibodies have, for example, beenproposed to target immune system cells to unwanted cells [U.S. Pat. No.4,676,980], and for treatment of HIV infection [WO 91/00360; WO92/200373; EP 03089]. It is contemplated that the antibodies may beprepared in vitro using known methods in synthetic protein chemistry,including those involving crosslinking agents. For example, immunotoxinsmay be constructed using a disulfide exchange reaction or by forming athioether bond. Examples of suitable reagents for this purpose includeiminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, forexample, in U.S. Pat. No. 4,676,980.

[1570] 6. Effector Function Engineering

[1571] It may be desirable to modify the antibody of the invention withrespect to effector function, so as to enhance, e.g., the effectivenessof the antibody in treating cancer. For example, cysteine residue(s) maybe introduced into the Fc region, thereby allowing interchain disulfidebond formation in this region. The homodimeric antibody thus generatedmay have improved internalization capability and/or increasedcomplement-mediated cell killing and antibody-dependent cellularcytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191-1195(1992) and Shopes, J. Immunol. 148: 2918-2922 (1992). Homodimericantibodies with enhanced anti-tumor activity may also be prepared usingheterobifunctional cross-linkers as described in Wolff et al. CancerResearch, 53: 2560-2565 (1993). Alternatively, an antibody canbeengineered that has dual Fc regions and may thereby have enhancedcomplement lysis and ADCC capabilities. See Stevenson et al.,Anti-Cancer Drug Design, 3: 219-230 (1989).

[1572] 7. Immunoconjugates

[1573] The invention also pertains to immunoconjugates comprising anantibody conjugated to a cytotoxic agent such as a chemotherapeuticagent, toxin (e.g., an enzymatically active toxin of bacterial, fungal,plant, or animal origin, or fragments thereof), or a radioactive isotope(i.e., a radioconjugate).

[1574] Chemotherapeutic agents useful in the generation of suchimmunoconjugates have been described above. Enzymatically active toxinsand fragments thereof that can be used include diphtheria A chain,nonbinding active fragments of diphtheria toxin, exotoxin A chain (fromPseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain,alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phrytolacaamericana proteins (PAPI, PAPII, and PAP-S), momordica charantiainhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin,mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. Avariety of radionuclides are available for the production ofradioconjugated antibodies. Examples include ²¹²Bi, ¹³¹I, ¹³¹In, ⁹⁰Y,and ¹⁸⁶Re.

[1575] Conjugates of the antibody and cytotoxic agent are made using avariety of bifunctional protein-coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane(IT), bifunctional derivatives of imidoesters (such as dimethyladipimidate HCL), active esters (such as disuccinimidyl suberate),aldehydes (such as glutareldehyde), bis-azido compounds (such as bis(p-azidobenzoyl) hexanediamine), bis-diazoniurn derivatives (such asbis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such astolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as1,5difluoro-2,4 dinitrobenzene). For example, a ricin immunotoxin can beprepared as described in Vitetta et al., Science, 238: 1098 (1987).Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent forconjugation of radionucleotide to the antibody. See WO94/11026.

[1576] In another embodiment, the antibody may be conjugated to a“receptor” (such streptavidin) for utilization in tumor pretargetingwherein the antibody-receptor conjugate is administered to the patient,followed by removal of unbound conjugate from the circulation using aclearing agent and then administration of a “ligand” (e.g., avidin) thatis conjugated to a cytotoxic agent (e.g., a radionucleotide).

[1577] 8. Immunoliposomes

[1578] The antibodies disclosed herein may also be formulated asimmunoliposomes. Liposomes containing the antibody are prepared bymethods known in the art, such as described in Epstein et al., Proc.Natl. Acad. Sci. USA, 82: 3688 (1985); Hwang et al., Proc. Natl Acad.Sci. USA, 77: 4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545.Liposomes with enhanced circulation time are disclosed in U.S. Pat. No.5,013,556.

[1579] Particularly useful liposomes can be generated by thereverse-phase evaporation method with a lipid composition comprisingphosphatidylcholine, cholesterol, and PEG-derivatizedphosphatidylethanolamine (PEG-PE). Liposomes are extruded throughfilters of defined pore size to yield liposomes with the desireddiameter. Fab′ fragments of the antibody of the present invention can beconjugated to the liposomes as described in Martin et al ., J. Biol.Chem., 257: 286-288 (1982) via a disulfide-interchange reaction. Achemotherapeutic agent (such as Doxorubicin) is optionally containedwithin the liposome. See Gabizon et al., J. National Cancer Inst.,81(19):1484 (1989).

[1580] 9. Pharmaceutical Compositions of Antibodies

[1581] Antibodies specifically binding a PRO polypeptide identifiedherein, as well as other molecules identified by the screening assaysdisclosed hereinbefore, can be administered for the treatment of variousdisorders in the form of pharmaceutical compositions.

[1582] If the PRO polypeptide is intracellular and whole antibodies areused as inhibitors, internalizing antibodies are preferred. However,lipofections or liposomes can also be used to deliver the antibody, oran antibody fragment, into cells. Where antibody fragments are used, thesmallest inhibitory fragment that specifically binds to the bindingdomain of the target protein is preferred. For example, based upon thevariable- region sequences of an antibody, peptide molecules can bedesigned that retain the ability to bind the target protein sequence.Such peptides can be synthesized chemically and/or produced byrecombinant DNA technology. See, e.g., Marasco et al., Proc. Natl. Acad.Sci. USA, 90: 7889-7893 (1993). The formulation herein may also containmore than one active compound as necessary for the particular indicationbeing treated, preferably those with complementary activities that donot adversely affect each other. Alternatively, or in addition, thecomposition may comprise an agent that enhances its function, such as,for example, a cytotoxic agent, cytokine, chemotherapeutic agent, orgrowth-inhibitory agent. Such molecules are suitably present incombination in amounts that are effective for the purpose intended.

[1583] The active ingredients may also be entrapped in microcapsulesprepared, for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly-(methylmethacylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles, andnanocapsules) or in macroemulsions. Such techniques are disclosed inRemington's Pharmaceutical Sciences, supra.

[1584] The formulations to be used for in Wvo administration must besterile. This is readily accomplished by filtration through sterilefiltration membranes.

[1585] Sustained-release preparations may be prepared. Suitable examplesof sustained-release preparations include semipermeable matrices ofsolid hydrophobic polymers containing the antibody, which matrices arein the form of shaped articles, e.g., films, or microcapsules. Examplesof sustained-release matrices include polyesters, hydrogels (forexample, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)),polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acidand γ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate,degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (injectable microspheres composed of lactic acid-glycolic acidcopolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid.While polymers such as ethylene-vinyl acetate and lactic acid-glycolicacid enable release of molecules for over 100 days, certain hydrogelsrelease proteins for shorter time periods. When encapsulated antibodiesremain in the body for a long time, they may denature or aggregate as aresult of exposure to moisture at 37° C., resulting in a loss ofbiological activity and possible changes in immunogenicity. Rationalstrategies can be devised for stabilization depending on the mechanisminvolved. For example, if the aggregation mechanism is discovered to beintermolecular S-S bond formation through thiodisulfide interchange,stabilization may be achieved by modifying sulfhydryl residues,lyophllizing from acidic solutions, controlling moisture content, usingappropriate additives, and developing specific polymer matrixcompositions.

[1586] G. Uses for anti-PRO Antibodies

[1587] The anti-PRO antibodies of the invention have various utilities.For example, anti-PRO antibodies may be used in diagnostic assays forPRO, e.g., detecting its expression in specific cells, tissues, orserum. Various diagnostic assay techniques known in the art may be used,such as competitive binding assays, direct or indirect sandwich assaysand immunoprecipitation assays conducted in either heterogeneous orhomogeneous phases [Zola, Monoclonal Antibodies: A Manual of Techniques,CRC Press, Inc. (1987) pp. 147-1581. The antibodies used in thediagnostic assays can be labeled with a detectable moiety. Thedetectable moiety should be capable of producing, either directly orindirectly, a detectable signal. For example, the detectable moiety Maybe a radioisotope, such as ³H, ¹⁴C, ³²P, ³⁵S, or ¹²⁵I, a fluorescent orchemiluminescent compound, such as fluorescein isothiocyanate,rhodamine, or luciferin, or an enzyme, such as alkaline phosphatase,beta-galactosidase or horseradish peroxidase. Any method known in theart for conjugating the antibody to the detectable moiety may beemployed, including those methods described by Hunter et al., Nature,144:945 (1962); David et al., Biochemistry, 13:1014 (1974); Pain et al.,J. Immunol. Meth., 40:219 (1981); and Nygren, J. Histochem. andCytochem., 30:407 (1982).

[1588] Anti-PRO antibodies also are useful for the affinity purificationof PRO from recombinant cell culture or natural sources. In thisprocess, the antibodies against PRO are immobilized on a suitablesupport, such a Sephadex resin or filter paper, using methods well knownin the art. The immobilized antibody then is contacted with a samplecontaining the PRO to be purified, and thereafter the support is washedwith a suitable solvent that will remove substantially all the materialin the sample except the PRO, which is bound to the immobilizedantibody. Finally, the support is washed with another suitable solventthat will release the PRO from the antibody.

[1589] The following examples are offered for illustrative purposesonly, and are not intended to limit the scope of the present inventionin any way.

[1590] All patent and literature references cited in the presentspecification are hereby incorporated by reference in their entirety.

EXAMPLES

[1591] Commercially available reagents referred to in the examples wereused according to manufacturer's instructions unless otherwiseindicated. The source of those cells identified in the followingexamples, and throughout the specification, by ATCC accession numbers isthe American Type Culture Collection, Rockville, Md.

Example 1 Extracellular Domain Homology Screening to Identify NovelPolypeptides and cDNA Encoding Therefor

[1592] The extracellular domain (ECD) sequences (including the secretionsignal sequence, if any) from about 950 known secreted proteins from theSwiss-Prot public database were used to search EST databases. The ESTdatabases included public databases (e.g., Dayhoff, GenBank), andproprietary databases (e.g. LIFESEQ™, Incyte Pharmaceuticals, Palo Alto,Calif.). The search was performed using the computer program BLAST orBLAST2 (Altschul and Gish, Methods in Enzymology 266: 460-480 (1996)) asa comparison of the ECD protein sequences to a 6 frame translation ofthe EST sequences. Those comparisons with a Blast score of 70 (or insome cases 90) or greater that did not encode known proteins wereclustered and assembled into consensus DNA sequences with the program“phrap” (Phil Green, University of Washington, Seattle, Wash.).

[1593] Using this extracellular domain homology screen, consensus DNAsequences were assembled relative to the other identified EST sequencesusing phrap. In addition, the consensus DNA sequences obtained wereoften (but not always) extended using repeated cycles of BLAST and phrapto extend the consensus sequence as far as possible using the sources ofEST sequences discussed above.

[1594] Based upon the consensus sequences obtained as described above,oligonucleotides were then synthesized and used to identify by PCR acDNA library that contained the sequence of interest and for use asprobes to isolate a clone of the full-length coding sequence for a PROpolypeptide. Forward (.f) and reverse (.r) PCR primers generally rangefrom 20 to 30 nucleotides and are often designed to give a PCR productof about 100-1000 bp in length. The probe (.p) sequences are typically40-55 bp in length. In some cases, additional oligonucleotides aresynthesized when the consensus sequence is greater than about 1-1.5 kbp.In order to screen several libraries for a full-length clone, DNA fromthe libraries was screened by PCR amplification, as per Ausubel et al.,Current Protocols in Molecular Biology, with the PCR primer pair. Apositive library was then used to isolate clones encoding the gene ofinterest using the probe oligonucleotide and one of the primer pairs.

[1595] The cDNA libraries used to isolate the cDNA clones wereconstructed by standard methods using commercially available reagentssuch as those from Invitrogen, San Diego, Calif. The cDNA was primedwith oligo dT containing a NotI site, linked with blunt to SalIhemikased adaptors, cleaved with NotI, sized appropriately by gelelectrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the Sfil site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

Example 2 Isolation of cDNA clones by Amylase Screening

[1596] 1. Preparation of Olio dT Primed cDNA Library

[1597] mRNA was isolated from a human tissue of interest using reagentsand protocols from Invitrogen, San Diego, Calif. (Fast Track 2). ThisRNA was used to generate an oligo dT prmed cDNA library in the vectorpRK5D using reagents and protocols from Life Technologies, Gaithersburg,Md. (Super Script Plasmid System). In this procedure, the doublestranded cDNA was sized to greater than 1000 bp and the SalI/NotIlinkered cDNA was cloned into XhoI/NotI cleaved vector. pRK5D is acloning vector that has an sp6 transcription initiation site followed byan SfiI restriction enzyme site preceding the XhoI/NotI cDNA cloningsites.

[1598] 2. Preparation of Random Primed cDNA Library

[1599] A secondary cDNA library was generated in order to preferentiallyrepresent the 5′ ends of the primary cDNA clones. Sp6 RNA was generatedfrom the primary library (described above), and this RNA was used togenerate a random primed cDNA library in the vector pSST-AMY.0 usingreagents and protocols from Life Technologies (Super Script PlasmidSystem, referenced above). In this procedure the double stranded cDNAwas sized to 500-1000 bp, Tinkered with blunt to NotI adaptors, cleavedwith SfiI, and cloned into SfiI/NotI cleaved vector. pSST-AMY.0 is acloning vector that has a yeast alcohol dehydrogenase promoter precedingthe cDNA cloning sites and the mouse amylase sequence (the maturesequence without the secretion signal) followed by the yeast alcoholdehydrogenase terminator, after the cloning sites. Thus, cDNAs clonedinto this vector that are fused in frame with amylase sequence will leadto the secretion of amylase from appropriately transfected yeastcolonies.

[1600] 3. Transformation and Detection

[1601] DNA from the library described in paragraph 2 above was chilledon ice to which was added electrocompetent DH10B bacteria (LifeTechnologies, 20 ml). The bacteria and vector mixture was thenelectroporated as recommended by the manufacturer. Subsequently, SOCmedia (Life Technologies, 1 ml) was added and the mixture was incubatedat 37° C. for 30 minutes. The transformants were then plated onto 20standard 150 mm LB plates containing ampicillin and incubated for 16hours (37° C.). Positive colonies were scraped off the plates and theDNA was isolated from the bacterial pellet using standard protocols,e.g. CsCl-gradient. The purified DNA was then carried on to the yeastprotocols below.

[1602] The yeast methods were divided into three categories: (1)Transformation of yeast with the plasmid/cDNA combined vector; (2)Detection and isolation of yeast clones secreting amylase; and (3) PCRamplification of the insert directly from the yeast colony andpurification of the DNA for sequencing and further analysis.

[1603] The yeast strain used was HD56-5A (ATCC-90785). This strain hasthe following genotype: MAT alpha, ura3-52, leu2-3, leu2-112, his3-11,his3-15, MAL⁺, SUC⁺, GAL⁺. Preferably, yeast mutants can be employedthat have deficient post-translational pathways. Such mutants may havetranslocation deficient alleles in sec71, sec72, sec62, with truncatedsec71 being most preferred. Alternatively, antagonists (includingantisense nucleotides and/or ligands) which interfere with the normaloperation of these genes, other proteins implicated in this posttranslation pathway (e.g., SEC61p, SEC72p, SEC62p, SEC63p, TDJ1p orSSA1p4p) or the complex formation of these proteins may also bepreferably employed in combination with the amylase-expressing yeast.

[1604] Transformation was performed based on the protocol outlined byGietz et al., Nucl. Acid. Res., 20:1425 (1992). Transformed cells werethen inoculated from agar into YEPD complex media broth (100 ml) andgrown overnight at 30° C. The YEPD broth was prepared as described inKaiser et al., Methods in Yeast Genetics, Cold Spring Harbor Press, ColdSpring Harbor, N.Y., p. 207 (1994). The overnight culture was thendiluted to about 2×10⁶ cells/ml (approx. OD600=0.1) into fresh YEPDbroth (500 ml) and regrown to 1×10⁷ cells/ml (approx. OD600₌0.4-0.5).

[1605] The cells were then harvested and prepared for transformation bytransfer into GS3 rotor bottles in a Sorval GS3 rotor at 5,000 rpm for 5minutes, the supernatant discarded, and then resuspended into sterilewater, and centrifuged again in 50 ml falcon tubes at 3,500 rpm in aBeckman GS-6KR centrifuge. The supernatant was discarded and the cellswere subsequently washed with LiAc/TE (10 ml, 10 mM Tris-HCl, 1 MM EDTApH 7.5, 100 mM Li₂OOCCH₃), and resuspended into LiAc/TE (2.5 ml).

[1606] Transformation took place by mixing the prepared cells (100 μl)with freshly denatured single stranded salmon testes DNA (LofstrandLabs, Gaithersburg, Md.) and transforming DNA (1 μg, vol.<10 μl) inmicrofuge tubes. The nixture was mixed briefly by vortexing, then 40%PEG/TE (600 μl, 40% polyethylene glycol4000, 10 mM Tris-HCl, 1 mM EDTA,100 mM Li₂OOCCH₃, pH 7.5) was added. This mixture was gently mixed andincubated at 30° C. while agitating for 30 minutes. The cells were thenheat shocked at 42° C. for 15 minutes, and the reaction vesselcentrifuged in a microfuge at 12,000 rpm for 5-10 seconds, decanted andresuspended into TE (500 μl, 10 mM Tris-HCl, 1 mM EDTA pH 7.5) followedby recentrifugation. The cells were then diluted into TE (1 ml) andaliquots (200 μl) were spread onto the selective media previouslyprepared in 150 mm growth plates (VWR).

[1607] Alternatively, instead of multiple small reactions, thetransformation was performed using a single, large scale reaction,wherein reagent amounts were scaled up accordingly.

[1608] The selective media used was a synthetic complete dextrose agarlacking uracil (SCD-Ura) prepared as described in Kaiser et al., Methodsin Yeast Genetics, Cold Spring Harbor Press, Cold Spring Harbor, N.Y.,p. 208-210 (1994). Transformants were grown at 30° C. for 2-3 days.

[1609] The detection of colonies secreting amylase was performed byincluding red starch in the selective growth media. Starch was coupledto the red dye (Reactive Red-120, Sigma) as per the procedure describedby Biely et al., Anal. Biochem., 172:176-179 (1988). The coupled starchwas incorporated into the SCD-Ura agar plates at a final concentrationof 0.15% (w/v), and was buffered with potassium phosphate to a pH of 7.0(50-100 mM final concentration).

[1610] The positive colonies were picked and streaked across freshselective media (onto 150 mm plates) in order to obtain well isolatedand identifiable single colonies. Well isolated single colonies positivefor amylase secretion were detected by direct incorporation of redstarch into buffered SCD-Ura agar. Positive colonies were determined bytheir ability to break down starch resulting in a clear halo around thepositive colony visualized directly.

[1611] 4. Isolation of DNA by PCR Amplification

[1612] When a positive colony was isolated, a portion of it was pickedby a toothpick and diluted into sterile water (30 μl) in a 96 wellplate. At this time, the positive colonies were either frozen and storedfor subsequent analysis or immediately amplified. An aliquot of cells(54 μl) was used as a template for the PCR reaction in a 25 μl volumecontaining: 0.5 μl Klentaq (Clontech, Palo Alto, Calif.); 4.0 μl 10 mMdNfP's (Perkin Elmer-Cetus); 2.5 μl Kentaq buffer (Clontech); 0.25 μlforward oligo 1; 0.25 μl reverse oligo 2; 12.5 μl distilled water. Thesequence of the forward oligonucleotide 1 was:

[1613] 5′-TGTAAAACGACGGCCAGT{overscore (TAAATAGACCTGCAATTATTAATCT)}-3′(SEQ ID NO:324)

[1614] The sequence of reverse oligonucleotide 2 was:

[1615] 5′-CAGGAAACAGCTATGACC{overscore (ACCTGCACACCTGCAAATCCATT)}-3′(SEQ ID NO:325)

[1616] PCR was then performed as follows: a. Denature 92° C.,  5 minutesb. 3 cycles of: Denature 92° C., 30 seconds Anneal 59° C., 30 secondsExtend 72° C., 60 seconds c. 3 cycles of: Denature 92° C., 30 secondsAnneal 57° C., 30 seconds Extend 72° C., 60 seconds d. 25 cycles of:Denature 92° C., 30 seconds Anneal 55° C., 30 seconds Extend 72° C., 60seconds e. Hold 4° C.

[1617] The underlined regions of the oligonucleotides annealed to theADH promoter region and the amylase region, respectively, and amplifieda 307 bp region from vector pSST-AMY.0 when no insert was present.Typically, the first 18 nucleotides of the 5′ end of theseoligonucleotides contained annealing sites for the sequencing primers.Thus, the total product of the PCR reaction from an empty vector was 343bp. However, signal sequence-fused cDNA resulted in considerably longernucleotide sequences.

[1618] Following the PCR, an aliquot of the reaction (5 μl) was examinedby agarose gel electrophoresis in a 1% agarose gel using aTris-Borate-EDTA (TBE) buffering system as described by Sambrook et al.,supra. Clones resulting in a single strong PCR product larger than 400bp were further analyzed by DNA sequencing after purification with a 96Qiaquick PCR clean-up column (Qiagen Inc., Chatsworth, Calif.).

Example 3 Isolation of cDNA Clones Encoding Human PRO213

[1619] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA28735. Based on the DNA28735consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO213.

[1620] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-TGGAGCAGCAATATGCCAGCC-3′ (SEQ ID NO:3) reverse PCRprimer 5′-TTTTCCACTCCTGTCGGGTTGG-3′ (SEQ ID NO:4)

[1621] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA28735 sequence which had the followingnucleotide sequence

[1622] hybridization probe

[1623] 5′-GGTGACACTTGCCAGTCAGATGTGGATGAATGCAGTGCTAGGAGGG-3′ (SEQ IDNO:5)

[1624] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO213 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal lungtissue.

[1625] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO213 herein designated as UNQ187(DNA30943-1163)] (SEQ ID NO:1) and the derived protein sequence forPRO213.

[1626] The entire nucleotide sequence of UNQ187 (DNA30943-1163) is shownin FIG. 1 (SEQ ID NO:1). Clone UNQ187 (DNA30943-1163) contains a singleopen reading frame with an apparent translational initiation site atnucleotide positions 336-338 and ending at the stop codon at nucleotidepositions 1221-1223 (FIG. 1). The predicted polypeptide precursor is 295amino acids long (FIG. 2). Clone UNQ187 (DNA30943-1163) has beendeposited with ATCC.

[1627] Analysis of the amino acid sequence of the full-length PRO213polypeptide suggests that a portion of it possesses significant homologyto the human growth arrest-specific gene 6 protein. More specifically,an analysis of the Dayhoff database (version 35.45 SwissProt 35)evidenced significant homology between the PRO213 amino acid sequenceand the following Dayhoff sequences, HSMHC3W5A_(—)6 and B48089.

Example 4 Isolation of cDNA Clones Encoding Human PRO274

[1628] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA36469. Based on the DNA36469consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO274. ESTs proprietary to Genentech were employed in the consensusassembly. The ESTs are shown in FIGS. 5-7 and are herein designatedDNA17873, DNA36157 and DNA28929, respectively.

[1629] Pairs of PCR primers (forward and reverse) were synthesized:forward PCR primer 1 (36469.f1) 5′-CTGATCCGGTTCTTGGTGCCCCTG-3′ (SEQ IDNO:11) forward PCR primer 2 (36469.f2) 5′-GCTCTGTCACTCACGCTC-3′ (SEQ IDNO:12) forward PCR primer 3 (36469.f3) 5′-TCATCTCTTCCCTCTCCC-3′ (SEQ IDNO:13) forward PCR primer 4 (36469.f4) 5′-CCTTCCGCCACGGAGTTC-3′ (SEQ IDNO:14) reverse PCR primer 1 (36469.r1) 5′-GGCAAAGTCCACTCCGATGATGTC-3′(SEQ ID NO:15) reverse PCR primer 2 (36469.r2)5′-GCCTGCTGTGGTCACAGGTCTCCG-3′ (SEQ ID NO:16)

[1630] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA36469 sequence which had the followingnucleotide sequence

[1631] hybridization probe (36469.p1)

[1632] 5′-TCGGGGAGCAGGCCTTGAACCGGGGCATTGCTGCTGTCAAGGAGG-3′ (SEQ IDNO:17)

[1633] In order to screen several libraries for a source of afuill-length clone, DNA from the libraries was screened by PCRampllfication with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO274 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB229).

[1634] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO274 [herein designated as UNQ241(DNA39987-1184)] (SEQ ID NO:1) and the derived protein sequence forPRO274.

[1635] The entire nucleotide sequence of UNQ241 (DNA39987-1184) is shownin FIG. 3 (SEQ ID NO:6). Clone UNQ241 (DNA39987-1184) contains a singleopen reading frame with an apparent translational initiation site atnucleotide positions 83-85 and ending at the stop codon at nucleotidepositions 1559-1561 (FIG. 3). The predicted polypeptide precursor is 492amino acids long (FIG. 4), has an estimated molecular weight of about54,241 daltons and an estimated pI of about 8.21. Clone UNQ241(DNA39987-1184) has been deposited with ATCC and is assigned ATCCdeposit no. 209786.

[1636] Analysis of the amino acid sequence of the full-length PRO274polypeptide suggests that it possesses significant homology to the Fn54protein. More specifically, an analysis of the Dayhoff database (version35.45 SwissProt 35) evidenced significant homology between the PRO274amino acid sequence and the following Dayhoff sequences, MMFN54S2_(—)1,MMFN54S1_(—)1, CELF48C1_(—)8, CEF38B7_(—)6, PRP3RAT, INK3_PIG,MTCY07A7_(—)13, YNAX_KLEAE, A47234 and HME2_MOUSE.

Example 5 Isolation of cDNA Clones Encoding Human PRO300

[1637] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA35930. Based on the DNA35930consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO300.

[1638] Forward and reverse PCR primers were synthesized: forward PCRprimer 1 (35930.f1) 5′-GCCGCCTCATCTTCACGTTCTTCC-3′ (SEQ ID NO:20)forward PCR primer 2 (35930.f2) 5′-TCATCCAGCTGGTGCTGCTC-3′ (SEQ IDNO:21) forward PCR primer 3 (35930.f3) 5′-CTTCTTCCACTTCTGCCTGG-3′ (SEQID NO:22) forward PCR primer 4 (35930.f4) 5′-CCTGGGCAAAAATGCAAC-3′ (SEQID NO:23) reverse PCR primer 1 (35930.r1) 5′-CAGGAATGTAGAAGGCACCCACGG-3′(SEQ ID NO:24) reverse PCR primer 2 (35930.r2)5′-TGGCACAGATCTTCACCCACACGG-3′ (SEQ ID NO:25)

[1639] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35930 sequence which had the followingnucleotide sequence

[1640] hybridization probe (35930.p1)

[1641] 5′-TGTCCATCATTATGCTGAGCCCGGGCGTGGAGAGTCAGCTCTACAAGCTG-3′ (SEQ IDNO:26)

[1642] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO300 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue.

[1643] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO300 [herein designated as UNQ263(DNA40625-1189)] (SEQ ID NO:18) and the derived protein sequence forPRO300.

[1644] The entire nucleotide sequence of UNQ263 (DNA40625-1189) is shownin FIG. 8 (SEQ ID NO:18). Clone UNQ263 (DNA40625-1189) contains a singleopen reading frame with an apparent translational initiation site atnucleotide positions 45-47 and ending at the stop codon at nucleotidepositions 1416-1418 (FIG. 8). The predicted polypeptide precursor is 457amino acids long (FIG. 9). Clone UNQ263 (DNA40625-1189) has beendeposited with ATCC and is assigned ATCC deposit no. 209788.

[1645] Analysis of the amino acid sequence of the full-length PRO300polypeptide suggests that portions of it possess significant homology tothe Diff 33 protein. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO300 amino acid sequence and the following Dayhoffsequence, HSU49188_(—)1.

Example 6 Isolation of cDNA Clones Encoding Human PRO284

[1646] Two cDNA sequences were isolated in the amylase screen describedin Example 2 and those cDNA sequences are herein designated DNA12982(see FIG. 12; human placenta-derived) and DNA15886 (see FIG. 13; humansalivary gland-derived). The DNA12982 and DNA15886 sequences were thenclustered and aligned, giving rise to a consensus nucleotide sequenceherein designated DNA18832.

[1647] Based on the DNA18832 consensus sequence, oligonucleotide probeswere generated and used to screen a human placenta library (LIB89)prepared as described in paragraph 1 of Example 2 above. The cloningvector was pRK5B (pRK5B is a precursor of pRK5D that does not containthe SfiI site; see, Holmes et al., Science, 253:1278-1280 (1991)), andthe cDNA size cut was less than 2800 bp.

[1648] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 (18832.est.f) 5′-TCGTACAGTTACGCTCTCCC-3′ (SEQ ID NO:31) forwardPCR primer 2 (18832.f) 5′-CTTGAGGAGCGTCAGAAGCG-3′ (SEQ ID NO:32) reversePCR primer (18832.r) 5′-ATAACGAATGAAGCCTCGTG-3′ (SEQ ID NO:33)

[1649] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA18832 sequence which had the followingnucleotide sequence

[1650] hybridization probe (18832.p)

[1651] 5′-GCTAATATCTGTAAGACGGCAGCTACAGCAGGCATCATTG-3′ (SEQ ID NO:34)

[1652] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pairs identified above. A positivelibrary was then used to isolate clones encoding the PRO284 gene usingthe probe oligonucleotide and one of the PCR primers.

[1653] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 167-169 and ending at the stop codon found atnucleotide positions 1022-1024 (FIG. 10; SEQ ID NO:27). The predictedpolypeptide precursoris 285 amino acids long, has a calculated molecularweight of approximately 32,190 daltons and an estimated pI ofapproximately 9.03. Analysis of the full-length PRO284 sequence shown inFIG. 11 (SEQ ID NO:28) evidences the presence of the following: a signalpeptide from about amino acid 1 to about amino acid 24, transmembranedomains from about amino acid 76 to about amino acid 96 and from aboutamino acid 171 to about amino acid 195 and a potential N-glycosylationsite from about amino acid 153 to about amino acid 156. Clone UNQ247(DNA23318-1211) has been deposited with ATCC on Apr. 21, 1998 and isassigned ATCC deposit no. 209787.

[1654] Analysis of the amino acid sequence of the full-length PRO284polypeptide suggests that it possesses no significant sequencesimilarity to any known protein. However, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced some degree of homologybetween the PRO284 amino acid sequence and the following Dayhoffsequences, JQ0124, CELE04A4_(—)5, AB006451_(—)1, AF030162_(—)1,IM23_YEAST, S71194, NIA_CUCMA, IM17_YEAST, I50479 and HUMZFHP_(—)1.

Example 7 Isolation of cDNA Clones Encoding Human PRO296

[1655] A cDNA sequence isolated in the amylase screen as described inExample 2 above was found, by BLAST and FastA sequence alignment, tohave sequence homology to a nucleotide sequence encoding sarcoma-associatedprotein SAS. This cDNA sequence is herein designated DNA23020(see FIG. 16). The DNA23020 sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ™,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.;http:/lbozeman.mbt.washington.edu/phrap.docs/phrap.html). The consensussequence obtained therefrom is herein designated DNA35858. Twoproprietary Genentech ESTs were employed in the assembly wherein thoseEST sequences are herein identified as DNA21971 (FIG. 17; SEQ ID NO:38)and DNA29037 (FIG. 18; SEQ ID NO:39).

[1656] Based on the DNA35858 consensus sequence, oligonucleotide probeswere generated and used to screen a human kidney library (LI]3228)library prepared as described in paragraph 1 of Example 2 above. Thecloning vector was pRK5B (pRK5B is a precursor of pRK5D that does notcontain the SfiI site; see, Holmes et al., Science, 253:1278-1280(1991)), and the cDNA size cut was less than 2800 bp.

[1657] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 (35858.f1) 5′-ACCCACGTCTGCGTTGCTGCC-3′ (SEQ ID NO:40) forwardPCR primer 2 (35858.f2) 5′-GAGAATATGCTGGAGAGG-3′ (SEQ ID NO:41) reversePCR primer (35858.r1) 5′-AGGAATGCACTAGGATTCGCGCGG-3′ (SEQ ID NO:42)

[1658] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35858 sequence which had the followingnucleotide sequence

[1659] hybridization probe (35858.p1)

[1660] 5′-GGCCCCAAAGGCAAGGACAAAGCAGCTGTCAGGGAACCTCCGCCG-3′ (SEQ IDNO:43)

[1661] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO296 gene usingthe probe oligonucleotide and one of the PCR primers.

[1662] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 174-176 and ending at the stop codon found atnucleotide positions 786-788 (FIG. 14; SEQ ID NO:35). The predictedpolypeptide precursor is 204 amino acids long, has a calculatedmolecular weight of approximately 22,147 daltons and an estimated p1 ofapproximately 8.37. Analysis of the full-length PRO296 sequence shown inFIG. 15 (SEQ ID NO:36) evidences the presence of the following: a signalpeptide from about amino acid 1 to about amino acid 34 and transmembranedomains from about amino acid 47 to about amino acid 63, from aboutamino acid 72 to about amino acid 95 and from about amino acid 162 toabout amino acid 182. Clone UNQ260 (DNA39979-1213) has been depositedwith ATCC on Apr. 21, 1998 and is assigned ATCC deposit no. 209789.

[1663] Analysis of the amino acid sequence of the full-length PRO296polypeptide suggests that it possesses significant sequence similarityto the sarcoma-amplified SAS protein, thereby indicating that PRO296 maybe a novel SAS homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO296 amino acid sequence and the following Dayhoffsequences, I58391, GEN11061, SSC2B04_(—)1, HSU81031_(—)2, CD63_RAT,CD63_MOUSE, CD63_HUMAN, AF022813_(—)1, CD63_RABIT and CO02_HUMAN.

Example 8 Isolation of cDNA Clones Encoding Human PRO329

[1664] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA35612. Based on the DNA35612consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO329.

[1665] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 (35612.f1) 5′-TGGGCTGTGTCCTCATGG-3′ (SEQ ID NO:46) forward PCRprimer 2 (35612.f2) 5′-TTTCCAGCGCCAATTCTC-3′ (SEQ ID NO:47) reverse PCRprimer 1 (35612.r1) 5′-AGTTCTTGGACTGTGATAGCCAC-3′ (SEQ ID NO:48) reversePCR primer 2 (35612.r2) 5′-AAACTTGGTTGTCCTCAGTGGCTG-3′ (SEQ ID NO:49)

[1666] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35612 sequence which had the followingnucleotide sequence

[1667] hybridization probe (35612.p1)

[1668] 5′-GTGAGGGACCTGTCTGCACTGAGGAGAGCAGCTGCCACACGGAGG-3′ (SEQ IDNO:50)

[1669] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pairs identified above. A positivelibrary was then used to isolate clones encoding the PRO329 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB6).

[1670] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO329 [herein designated as UNQ291(DNA40594-1233)] (SEQ ID NO:44) and the derived protein sequenceforPRO329.

[1671] The entire nucleotide sequence of UNQ291 (DNA40594-1233) is shownin FIG. 19 (SEQ ID NO:44). Clone UNQ291 (DNA40594-1233) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 9-11 and ending at the stop codon at nucleotidepositions 1086-1088 (FIG. 19). The predicted polypeptide precursor is359 amino acids long (FIG. 20). The fIlU-length PRO329 protein shown inFIG. 20 has an estimated molecular weight of about 38,899 daltons and apI of about 5.21. Clone UNQ291 (DNA40594-1233) has been deposited withATCC on Feb. 5, 1998 and is assigned ATCC deposit no. 209617.

[1672] Analysis of the amino acid sequence of the fullength PRO329polypeptide suggests that it possesses significant sequence similarityto a high affinity immunoglobulin F. receptor protein. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant homology between the PRO329 aminoacid sequence and the following Dayhoff sequences, FCG1_HUMAN,FCG0_HUMAN, P_R91439, P_R22549, P_R91438, P_W00859, P_R20811, P_R22550,HUMCD6406_(—)1 and FCG1_MOUSE.

Example 9 Isolation of cDNA Clones Encoding Human PRO362

[1673] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA42257. Based on the DNA42257consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO362.

[1674] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 (42257.f1) 5′-TATCCCTCCAATTGAGCACCCTGG-3′ (SEQ ID NO:53)forward PCR primer 2 (42257.f2) 5′-GTCGGAAGACATCCCAACAAG-3′ (SEQ IDNO:54) reverse PCR primer 1 (42257.r1) 5′-CTTCACAATGTCGCTGTGCTGCTC-3′(SEQ ID NO:55) reverse PCR primer 2 (42257.r2)5′-AGCCAAATCCAGCAGCTGGCTTAC-3′ (SEQ ID NO:56)

[1675] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA42257 sequence which had the followingnucleotide sequence

[1676] hybridization probe (42257.p1)

[1677] 5′-TGGATGACCGGAGCCACTACACGTGTGAAGTCACCTGGCAGACTCCTGAT-3′ (SEQ IDNO:57)

[1678] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pairs identified above. A positivelibrary was then used to isolate clones encoding the PRO362 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal braintissue (LIB153).

[1679] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO362 [herein designated as UNQ317(DNA45416-1251)] (SEQ ID NO:51) and the derived protein sequence forPRO362.

[1680] The entire nucleotide sequence of UNQ317 (DNA45416-1251) is shownin FIG. 21 (SEQ ID NO:51). Clone UNQ317 (DNA45416-1251) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 119-121 and ending at the stop codon atnucleotide positions 1082-1084 (FIG. 21). The predicted polypeptideprecursor is 321 amino acids long (FIG. 22). The full-length PRO362protein shown in FIG. 2 has an estimated molecular weight of about35,544 daltons and a pI of about 8.51. Analysis of the full-lengthPRO362 polypeptide as shown in FIG. 22 evidences the presence of aglycosaminoglycan attachment site at about amino acid 149 to about aminoacid 152 and a transmembrane domain from about amino acid 276 to aboutamino acid 306. Clone UNQ317 (DNA45416-1251) has been deposited withATCC on Feb. 5, 1998 and is assigned ATCC deposit no. 209620.

[1681] Analysis of the amino acid sequence of the full-length PRO362polypeptide suggests that it possesses significant sequence similarityto the A33 antigen protein and the HCAR protein. More specifically, ananalysis of the Dayhoff database (version 35.45 SwissProt 35) evidencedsignificant homology between the PRO362 amino acid sequence and thefollowing Dayhoff sequences, AB002341_(—)1, HSU55258_(—)1,HSC7NRCAM_(—)1, RNU81037_(—)1, A33_HUMAN, P_W14158, NMNCAMRI_(—)1,HSTITINN2_(—)1, S71824_(—)1 and HSU63041_(—)1.

Example 10 Isolation of cDNA Clones Encoding Human PRO363

[1682] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA42828. Based on the DNA42828consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO363.

[1683] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer (42828.f1) 5′-CCAGTGCACAGCAGGCAACGAAGC-3′ (SEQ IDNO:60) reverse PCR primer (42828.r1) 5′-ACTAGGCTGTATGCCTGGGTGGGC-3′ (SEQID NO:61)

[1684] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA42828 sequence which had the followingnucleotide sequence

[1685] hybridization probe (42828.p1)

[1686] 5′-GTATGTACAAAGCATCGGCATGGTTGCAGGAGCAGTGACAGGC-3′ (SEQ ID NO:62)

[1687] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO363 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LB227).

[1688] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO363 [herein designated as UNQ318(DNA45419-1252)] (SEQ ID NO:58) and the derived protein sequence forPRO363.

[1689] The entire nucleotide sequence of UNQ318 (DNA45419-1252) is shownin FIG. 23 (SEQ ID NO:58). Clone UNQ318 (DNA45419-1252) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 190-192 and ending at the stop codon atnucleotide positions 1309-1311 (FIG. 23). The predicted polypeptideprecursor is 373 amino acids long (FIG. 24). The full-length PRO363protein shown in FIG. 24 has an estimated molecular weight of about41,281 daltons and a pI of about 8.33. A transmembrane domain exists atamino acids 221 to 254 of the amino acid sequence shown in FIG. 24 (SEQID NO:59). The PRO363 polypeptide also possesses at least two myelin P0protein domains from about amino acids 15 to 56 and from about aminoacids 87 to 116. Clone UNQ318 (DNA45419-1252) has been deposited withATCC on Feb. 5, 1998 and is assigned ATCC deposit no. 209616.

[1690] Analysis of the amino acid sequence of the full-length PRO363polypeptide suggests that it possesses significant sequence similarityto the cell surface protein HCAR, thereby indicating that PRO363 may bea novel HCAR homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO363 amino acid sequence and the following Dayhoffsequences, HS46KDA_(—)1, HSU90716_(—)1, MMCARH_(—)1, MMCARHOM_(—)1,MMU90715_(—)1, A33_HUMAN, P_W14146, P_W14158, A42632 and B42632.

Example 11 Isolation of cDNA Clones Encoding Human PRO868

[1691] A consensus sequence was obtained relative to a variety of ESTsequences as described in Examnple I above, wherein the consensussequence obtained is herein designated DNA38133. Based on the DNA38133consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO868.

[1692] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer (38133.f1) 5′-GTAGCAGTGCACATGGGGTGTTGG-3′ (SEQ IDNO:65) reverse PCR primer (38133.r1) 5′-ACCGCACATCCTCAGTCTCTGTCC-3′ (SEQID NO:66)

[1693] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA38133 sequence which had the followingnucleotide sequence

[1694] hybridization probe (38133.p1)

[1695] 5′-ACGATGATCGCGGGCTCCCTTCTCCTGCTTGGATTCCTTAGCACCACCAC-3′ (SEQ IDNO:67)

[1696] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO868 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1697] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO868 [herein designated as UNQ437(DNA52594-1270)] (SEQ ID NO:63) and the derived protein sequence forPRO868.

[1698] The entire nucleotide sequence of UNQ437 (DNA52594-1270) is shownin FIG. 25 (SEQ ID NO:63). Clone UNQ437 (DNA52594-1270) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 325-327 and ending at the stop codon atnucleotide positions 2290-2292 FIG. 25). The predicted polypeptideprecursor is 655 amino acids long (FIG. 26). The full-length PRO868protein shown in FIG. 26 has an estimated molecular weight of about71,845 daltons and apI of about 8.22. Analysis of the full-length PRO868polypeptide sequence demonstrates the presence of conservedcysteine-containing domains from about amino acid 66 to about amino acid78 and from about amino acid 123 to about amino acid 134 of the sequenceshown in FIG. 26 (SEQ ID NO:3), a TNFR death domain from about aminoacid 85 to about amino acid 110, a FASA_mouse death domain block fromabout amino acid 159 to about amino acid 175 and a transmembrane domainfrom about amino acid 347 to about amino acid 375. Clone UNQ437(DNA52594-1270) has been deposited with ATCC on Mar. 17, 1998 and isassigned ATCC deposit no. 209679 Analysis of the amino acid sequence ofthe full-length PRO868 polypeptide suggests that it possessessignificant sequence similarity to the tumor necrosis factor receptorprotein, thereby indicating that PRO868 may be a novel member of thetumor necrosis factor receptor family. More specifically, an analysis ofthe Dayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO868 amino acid sequence and the followingDayhoff sequences, RNU94330_(—)1, P_R99933, PR_R99945, P_R99950,HSU94332_(—)1, CD40_HUMAN, S63368_(—)1, TNR2_HUMAN, MVU87844_(—)1 ANDCVU87837_(—)1.

Example 12 Isolation of cDNA Clones Encoding Human PRO382

[1699] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA30892. Based on the DNA30892consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO382.

[1700] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-TGACATCGCCCTTATGAAGCTGGC-3′ (SEQ ID NO:70) reversePCR primer 5′-TACACGTCCCTGTGGTTGCAGATC-3′ (SEQ ID NO:71)

[1701] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA30892 sequence which had the followingnucleotide sequence

[1702] hybridization probe

[1703] 5′-CGTTCAATGCAGAAATGATCCAGCCTGTGTGCCTGCCCAACTCTGAAGAG-3′ (SEQ IDNO:72)

[1704] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO382 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1705] DNA sequencing of the clones isolated as described above gave thefll-length DNA sequence for PRO382 [herein designated as UNQ323(DNA45234-1277)] (SEQ ID NO:68) and the derived protein sequence forPRO382.

[1706] The entire nucleotide sequence of UNQ323 (DNA45234-1277) is shownin FIG. 27 (SEQ ID NO:68). Clone UNQ323 (DNA45234-1277) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 126-128 and ending at the stop codon atnucleotide positions 1485-1487 (FIG. 27). The predicted polypeptideprecursor is 453 amino acids long (FIG. 28). The full-length PRO382protein shown in FIG. 28 has an estimated molecular weight of about49,334 daltons and a pI of about 6.32. Analysis of the native PRO382amino acid sequence shown in FIG. 28 (SEQ ID NO:69) indicates thepresence of a putative transmembrane domain from about amino acid 240 toabout amino acid 284, a putative signal peptide at about amino acid 1 toabout amino acid 20, a putative apple domain at about amino acid 386 toabout amino acid 419, a putative Kringle domain at about amino acid 394to about amino acid 406 and a histidine-containing protease active siteat about amino acid 253 to about amino acid 258. Clone UNQ323(DNA45234-1277) has been deposited with ATCC on Mar. 5, 1998 and isassigned ATCC deposit no. 209654.

[1707] Analysis of the amino acid sequence of the flil-length PRO382polypeptide suggests that it possess significanthomology to serineprotease proteins, thereby indicating that PRO382 may be anovel serineprotease. Specifically, an analysis of the Dayhoff database (version35.45 SwissProt 35) evidenced significant homology between the PRO382amino acid sequence and the following Dayhoff sequences, HSU75329_(—)1,ENTK_MOUSE, HEPS_HUMAN, AF030065_(—)1, HEPS_RAT, PLMN_PIG, P_R89430,P_R89435, PLMN_HORSE, PLMN_BOVIN and P_R83959.

Example 13 Isolation of cDNA Clones Encoding Human PRO545

[1708] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA44706. An EST proprietary toGenentech was employed in the consensus assembly and is hereindesignated DNA13217 (FIG. 31; SEQ ID NO:75). Based on the DNA44706consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO545.

[1709] Forward and reverse PCR primers were synthesized: forward PCRprimer 1 5′-GTCTCAGCACGTGTTCTGGTCTCAGGG-3′ (SEQ ID NO:76) forward PCRprimer 2 5′-CATGAGCATGTGCACGGC-3′ (SEQ ID NO:77) forward PCR primer 35′-TACCTGCACGATGGGCAC-3′ (SEQ ID NO:78) forward PCR primer 45′-CACTGGGCACCTCCCTTC-3′ (SEQ ID NO:79) reverse PCR primer 15′-CTCCAGGCTGGTCTCCAAGTCCTTCC-3′ (SEQ ID NO:80) reverse PCR primer 25′-TCCCTGTTGGACTCTGCAGCTTCC-3′ (SEQ ID NO:81) reverse PCR primer 35′-CTTCGCTGGGAAGAGTTTG-3′ (SEQ ID NO:82)

[1710] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA44706 sequence which had the followingnucleotide sequence

[1711] hybridization probe

[1712] 5′-GTGCAACCAACAGATACAAACTCTTCCCAGCGAAGAAGCTGAAAAGCGTC-3′ (SEQ IDNO:83)

[1713] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO545gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human placentatissue (LIB90).

[1714] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO545 [herein designated as UNQ346(DNA49624-1279)] (SEQ ID NO:73) and the derived protein sequence forPRO545.

[1715] The entire nucleotide sequence of UNQ346 (DNA49624-1279) is shownin FIG. 29 (SEQ ID NO:73). Clone UNQ346 (DNA49624-1279) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 311-313 and ending at the stop codon atnucleotide positions 2516-2518 (FIG. 29). The predicted polypeptideprecursor is 735 amino acids long (FIG. 30). The full-length PRO545protein shown in FIG. 30 has an estimated molecular weight of about80,177 daltons and a pI of about 7.08. Important regions of the PRO545amino acid sequence include the signal peptide, corresponding to aminoacids 1-28, five potential N-glycosylation sites, from about amino acid111-114, amino acids 146-149, amino acids 348-351, amino acids 449-452,and amino acids 648-651, and a neutral zinc metallopeptidase,zinc-binding region signature sequence, from about amino acids 344-353.Clone UNQ346 (DNA49624-1279) has been deposited with ATCC and isassigned ATCC deposit no. 209655.

Example 14 Isolation of cDNA Clones Encoding Human PRO617

[1716] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA42798. Based on the DNA42798sequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence for PRO617.

[1717] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-ACGGGCACACTGGATCCCAAATG-3′ (SEQ ID NO:86) reversePCR primer 5′-GGTAGAGATGTAGAAGGGCAAGCAAGACC-3′ (SEQ ID NO:87)

[1718] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA42798 sequence which had the followingnucleotide sequence

[1719] hybridization probe

[1720] 5′-GCTCCCTACCCGTGCAGGTTTCTTCATTTGTTCCTTTAACCAGTATGCCG-3′ (SEQ IDNO:88)

[1721] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO617 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1722] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO617 [herein designated as UNQ353(DNA48309-1280)] (SEQ ID NO:1) and the derived protein sequence forPRO617.

[1723] The entire nucleotide sequence of UNQ353 (DNA48309-1280) is shownin FIG. 32 (SEQ ID NO:84). Clone UNQ353 (DNA48309-1280) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 723-725 and ending at the stop codon atnucleotide positions 924-926 (FIG. 32). The predicted polypeptideprecursor is 67 amino acids long (FIG. 33). The full-length PRO617protein shown in FIG. 33 has an estimated molecular weight of about6,981 daltons and a pI of about 7.47. Analysis of the PRO617 amino acidsequence also evidences the existence of a putative signal peptide fromabout amino acid 15 to about amino acid 27 and a putative protein kinaseC phosphorylation site from about amino acid 41 to about amino acid 43.Clone UNQ353 (DNA48309-1280) has been deposited on Mar. 5, 1998 withATCC and is assigned ATCC deposit no. 209656.

[1724] Analysis of the amino acid sequence of the full-length PRO617polypeptide suggests that it possesses significant homology to the CD24protein, thereby indicating that PRO617 may be a novel CD24 homolog.More specifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant homology between the PRO617 aminoacid sequence and the following Dayhoff sequences, CD24_HUMAN,CD24_MOUSE, S15785, CD24_RAT, VGE BPG4, MSE5_HUMAN, HSMHC3W36A_(—)2,MLU15184_(—)8, P R85075, SEPL_HUMAN and MTCY63_(—)13.

Example 15 Isolation of cDNA Clones Encoding Human PRO700

[1725] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA30837. Based on the DNA30837consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO700.

[1726] Forward and reverse PCR primers were synthesized: forward PCRprimer 1 5′-ATGTTCTTCGCGCCCTGGTG-3′ (SEQ ID NO:91) forward PCR primer 25′-CCAAGCCAACACACTCTACAG-3′ (SEQ ID NO:92) reverse PCR primer 15′-AAGTGGTCGCCTTGTGCAACGTGC-3′ (SEQ ID NO:93) reverse PCR primer 25′-GGTCAAAGGGGATATATCGCCAC-3′ (SEQ ID NO:94)

[1727] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA30837 sequence which had the followingnucleotide sequence

[1728] hybridization probe

[1729] 5′-GCATGGAAGATGCCAAAGTCTATGTGGCTAAAGTGGACTGCACGGCCCA-3′ (SEQ IDNO:95)

[1730] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO700gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1731] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO700 [herein designated as UNQ364(DNA46776-1284)] (SEQ ID NO:89) and the derived protein sequence forPRO700.

[1732] The entire nucleotide sequence of UNQ364 (DNA46776-1284) is shownin FIG. 34 (SEQ ID NO:89). Clone UNQ364 (DNA46776-1284) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 33-35 and ending at the stop codon at nucleotidepositions 1329-1331 (FIG. 34). The predicted polypeptide precursor is432 amino acids long (FIG. 35). The full-length PRO700 protein shown inFIG. 35 has an estimated molecular weight of about 47,629 daltons andapI of about 5.90. Important regions of the amino acid sequence ofPRO700 include the signal peptide, corresponding to amino acids fromabout 1 to 33, regions homologous to disulfide isomerase, correspondingto amino acids from about 82-99, 210-255, and 345-360, a tyrosine kinasephosphorylation site, corresponding to amino acids from about 143-151,and an endoplasmic reticulum targeting sequence, corresponding to aminoacids from about 429-432. Clone UNQ364 (DNA46776-1284) has beendeposited with ATCC and is assigned ATCC Deposit No. 209721.

Example 16 Isolation of cDNA Clones Encoding Human PRO702

[1733] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA36623. Based on the DNA36623consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO702.

[1734] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer (36623.f1) 5′-CGCTGACTATGTTGCCAAGAGTGG-3′ (SEQ IDNO:98) reverse PCR primer (36623.r1) 5′-GATGATGGAGGCTCCATACCTCAG-3′ (SEQID NO:99)

[1735] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA36623 sequence which had the followingnucleotide sequence

[1736] hybridization probe (36623.p1)

[1737] 5′-GTGTTCATTGGCGTGAATGACCTTGAAAGGGAGGGACAGTACATGTTCAC-3′ (SEQ IDNO:100)

[1738] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO702 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB229).

[1739] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO702 [herein designated as UNQ366(DNA50980-1286)] (SEQ ID NO:96) and the derived protein sequence forPRO702.

[1740] The entire nucleotide sequence of UNQ366 (DNA50980-1286) is shownin FIG. 36 (SEQ ID NO:96). Clone UNQ366 (DNA50980-1286) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 22-24 and ending at the stop codon at nucleotidepositions 853-855 (FIG. 36). The predicted polypeptide precursor is 277amino acids long (FIG. 37). The full-length PRO702 protein shown in FIG.37 has an estimated molecular weight of about 30,645 daltons and a pI ofabout 7.47. Analysis of the full-length native PRO702 amino acidsequence evidences the presence of a putative signal peptide from aboutamino acid 1 to about amino acid 25, potential N-glycosylation sitesfrom about amino acid 230 to about amino acid 233 and from about aminoacid 258 to about amino acid 261 and a C-type lectin domain signaturesequence from about amino acid 248 to about amino acid 270. Clone UNQ366(DNA50980-1286) has been deposited with ATCC on Mar. 31, 1998 and isassigned ATCC deposit no. 209717.

[1741] Analysis of the amino acid sequence of the full-length PRO702polypeptide suggests that it possesses significant sequence similarityto the conglutinin protein, thereby indicating that PRO702 may be anovel conglutinin homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO702 amino acid sequence and the following Dayhoffsequences, S32436, P_R75642, P_W18780, P_W18781, A53330, AC002528_(—)1,HSPPA2IC0_(—)1, CA21_HUMAN, CA14_HUMAN and A61262.

Example 17 Isolation of cDNA Clones Encoding Human PRO703

[1742] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43047. Based on the DNA43047consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO703.

[1743] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-GAGAGCCATGGGGCTCCACCTG-3′ (SEQ ID NO:103) reverse PCR primer 15′-GGAGAATGTGGCCACAAC-3′ (SEQ ID NO:104) reverse PCR primer 25′-GCCCTGGCACAGTGACTCCATAGACG-3′ (SEQ ID NO:105) reverse PCR primer 35′-ATCCACTTCAGCGGACAC-3′ (SEQ ID NO:106)

[1744] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA40654 sequence which had the followingnucleotide sequence

[1745] hybridization probe

[1746] 5′-CCAGTGCCAGGATACCTCTCTTCCCCCCAGAGCATAACAGACACG-3′ (SEQ IDNO:107)

[1747] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO703gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1748] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO703 [herein designated as UNQ367(DNA50913-1287)] (SEQ ID NO:101) and the derived protein sequence forPRO703.

[1749] The entire nucleotide sequence of UNQ367 (DNA50913-1287) is shownin FIG. 38 (SEQ ID NO:101). Clone UNQ367 (DNA50913-1287) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 115-117 and ending at the stop codon atnucleotide positions 2305-2307 (FIG. 38). The predicted polypeptideprecursor is 730 amino acids long (FIG. 39). The full-length PRO703protein shown in FIG. 39 has an estimated molecular weight of about78,644 daltons, and a pI of about: 7.65. Important regions of the PRO703amino acid sequence include the signal peptide, a cAMP- andcGMP-dependent protein kinase phosphorylation site, a CUB domain proteinmotif, N-glycosylation sites and a putative AMP-binding domainsignature. Clone UNQ367 (DNA50913-1287) has been deposited with ATCC andis assigned ATCC deposit no. 209716.

Example 18 Isolation of cDNA Clones Encoding Human PRO705

[1750] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43437. Based on the DNA43437consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the fuillength coding sequence forPRO705.

[1751] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-AAGCGTGACAGCGGGCACGTC-3′ (SEQ ID NO:110) reversePCR primer 5′-TGCACAGTCTCTGCAGTGCCCAGG-3′ (SEQ ID NO:111)

[1752] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA43437 sequence which had the followingnucleotide sequence

[1753] hybridization probe (43437.p1)

[1754] 5′-GAATGCTGGAACGGGCACAGCAAAGCCAGATACTTGCCTG-3′ (SEQ ID NO:112)

[1755] In order to screen several libraries for a source of a fu-lengthclone, DNA from the libraries was screened by PCR amplification with thePCR primer pair identified above. A positive library was then used toisolate clones encoding the PRO705 gene using the probe oligonucleotideand one of the PCR primers. RNA for construction of the cDNA librarieswas isolated from human fetal kidney tissue (LIB227).

[1756] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO705 [herein designated as UNQ369(DNA50914-1289)] (SEQ ID NO:108) and the derived protein sequence forPRO705.

[1757] The entire nucleotide sequence of UNQ369 (DNA50914-1289) is shownin FIG. 40 (SEQ ID NO:108). Clone UNQ369 (DNA50914-1289) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 566-568 and ending at the stop codon atnucleotide positions 2231-2233 (FIG. 40). The predicted polypeptideprecursor is 555 amino acids long (FIG. 41). The full-length PRO705protein shown in FIG. 41 has an estimated molecular weight of about62,736 daltons and a pI of about 5.36. Analysis of the full-lengthPRO705 sequence as shown in FIG. 41 evidences the presence of thefollowing: a signal peptide from about amino acid 1 to about amino acid23, a eukaryotic DNA topoisomerase 1 active site from about amino acid418 to about amino acid 436, and various regions that show homology tovarious glypican proteins from about amino acid 237 to about amino acid279, about amino acid 421 to about amino acid 458, about amino acid 53to about amino acid 74, about amino acid 466 to about amino acid 504,about amino acid 308 to about amino acid 355, about amino acid 104 toabout amino acid 156 and about amino acid 379 to about amino acid 410.Clone UNQ369 (DNA50914-1289) has been deposited with ATCC on Mar. 31,1998 and is assigned ATCC deposit no.209722.

[1758] Analysis of the amino acid sequence of the full-length PRO705polypeptide suggests that it possesses significant sequence similarityto the K-glypican protein, thereby indicating that PRO705 may be a novelglypican protein family member. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO705 amino acid sequence and the followingDayhoff sequences, GPCK_MOUSE, GLYP_CHICK, GLYP_RAT, GLYP_HUMAN,GPC2_RAT, GPC5_HUMAN, GPC3_HUMAN, GPC3_RAT, P_(—R)30168 andCEC03H12_(—)2.

Example 19 Isolation of cDNA Clones Encoding Human PRO708

[1759] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA34024. Based on the DNA34024consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO708.

[1760] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CCCAACCCAACTGTTTACCTCTGG-3′ (SEQ ID NO:115)reverse PCR primer 5′-CTCTCTGAGTGTACATCTGTGTGG-3′ (SEQ ID NO:116)

[1761] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA34024 sequence which had the followingnucleotide sequence

[1762] hybridization probe

[1763] 5′-GCCACCCTACCTCAGAAACTGAAGGAGG NTATTCAACGCATATGGTCGG-3′ (SEQ IDNO:117)

[1764] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO708 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrowtissue (LIB255).

[1765] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO708 herein designated as UNQ372(DNA48296-1292)] (SEQ ID NO:113) and the derived protein sequence forPRO708.

[1766] The entire nucleotide sequence of UNQ372 (DNA48296-1292) is shownin FIGS. 42A-B (SEQ ID NO:113). Clone UNQ372 (DNA48296-1292) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 891-893 and ending at the stop codon atnucleotide positions 2436-2438 (FIGS. 42A-B). The predicted polypeptideprecursor is 515 amino acids long (FIG. 43). The full-length PRO708protein shown in FIG. 43 has an estimated molecular weight of about56,885 daltons and a pI of about 6.49. Analysis of the PRO708 amino acidsequence shown in FIG. 43 (SEQ ID NO:114) evidences the existence of aputative signal peptide at about amino acid 1 to about amino acid 37,putative sulfatase signature sequences at about amino acid 120 to aboutamino acid 132 and about amino acid 168 to about amino acid 177, aputative tyrosine kinase phosphorylation site from about amino acid 163to about amino acid 169 and potential N-glycosylation sites from aboutamino acid 157 to about amino acid 160, about amino acid 306 to aboutamino acid 309 and about amino acid 318 to about amino acid 321. CloneUNQ372 (DNA48296-1292) has been deposited with ATCC on Mar. 11, 1998 andis assigned ATCC deposit no. 209668.

[1767] Analysis of the amino acid sequence of the fti31-length PRO708polypeptide suggests that it possesses significant homology to the arylsulfatase proteins, thereby indicating that PRO708 may be a novel arylsulfatase homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO708 amino acid sequence and the following Dayhoffsequences, ARSB_HUMAN, CELC54D2_(—)2, G02857, STS_HUMAN, I37186, I37187,GEN12648, CELD1014_(—)7, GA6S_HUMAN and SPHM_HUMAN.

Example 20 Isolation of cDNA Clones Encoding Human PRO320

[1768] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA28739. Based on the DNA28739consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO320.

[1769] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CCTCAGTGGCCACATGCTCATG-3′ (SEQ ID NO:120) reversePCR primer 5′-GGCTGCACGTATGGCTATCCATAG-3′ (SEQ ID NO:121)

[1770] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA28739 sequence which had the followingnucleotide sequence

[1771] hybridization probe

[1772] 5′-GATAAACTGTCAGTACAGCTGTGAAGACACAGAAGAAGGGCCACAGTGCC-3′ (SEQ IDNO:122)

[1773] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO320 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal lungtissue (LIB25).

[1774] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO320 [herein designated as UNQ281(DNA32284-1307)] (SEQ ID NO:118) and the derived protein sequence forPRO320.

[1775] The entire nucleotide sequence of UNQ281 (DNA32284-1307) is shownin FIG. 44 (SEQ ID NO:118). Clone UNQ281 (DNA32284-1307) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 135-137 and ending at the stop codon atnucleotide positions 1149-1151 (FIG. 44). The predicted polypeptideprecursor is 338 amino acids long (FIG. 45). The full-length PRO320protein shown in FIG. 45 has an estimated molecular weight of about37,143 daltons and a pI of about 8.92. Important regions of the PRO320amino acid sequence include the signal peptide, corresponding to aminoacids 1-21, an EGF-like domain cysteine pattern signature, correspondingto amino acids 80-91, and three calcium- binding EGF-like domains,corresponding to amino acids 103-124, 230-151 and 185-206, respectively.Clone UNQ281 (DNA32284-1307) has been deposited with ATCC and isassigned ATCC deposit no. 209670.

Example 21 Isolation of cDNA Clones Encoding Human PRO324

[1776] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA34347. Based on the DNA34347consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO324.

[1777] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 5′-GCAATGAACTGGGAGCTGC-3′ (SEQ ID NO:125) forward PCR primer 25′-CTGTGAATAGCATCCTGGG-3′ (SEQ ID NO:126) forward PCR primer 35′-CTTTTCAAGCCACTGGAGGG-3′ (SEQ ID NO:127) reverse PCR primer 15′-CTGTAGACATCCAAGCTGGTATCC-3′ (SEQ ID NO:128) reverse PCR primer 25′-AAGAGTCTGCATCCACACCACTC-3′ (SEQ ID NO:129)

[1778] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA34347 sequence which had the followingnucleotide sequence

[1779] hybridization probe

[1780] 5′-ACCTGACGCTACTATGGGCCGAGTGGCAGGGACGACGCCCAGAATG-3′ (SEQ IDNO:130)

[1781] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO324gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB6).

[1782] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO324 [herein designated as UNQ285(DNA36343-1310)] (SEQ ID NO:123) and the derived protein sequence forPRO324.

[1783] The entire nucleotide sequence of UNQ285 (DNA36343-1310) is shownin FIG. 46 (SEQ ID NO:123). Clone UNQ285 (DNA36343-1310) containsasingle open reading frame with an apparent translational initiationsite at nucleotide positions 144-146 and ending at the stop codon atnucleotide positions 1011-1013 (FIG. 46). The predicted polypeptideprecursor is 289 amino acids long (FIG. 47). The full-length PRO324protein shown in FIG. 47 has an estimated molecular weight of about32,268 daltons and a pI of about 9.21. Analysis of the PRO324polypeptide sequence shown in FIG. 47 (SEQ ID NO:124) evidence thepresence of the following: a signal peptide from about amino acid 1 toabout amino acid 31, a transmembrane domain from about amino acid 136 toabout amino acid 157, tyrosine kinase phosphorylation sites from aboutamino acid 106 or about amino acid 107 to about amino acid 113 andregions that are homologous to short-chain alcohol dehydrogenase regionsfrom about amino acid 80 to about amino acid 90, from about amino acid131 to about amino acid 168, from about amino acid 1 to about amino acid13 and from about amino acid 176 to about amino acid 185. Clone UNQ285(DNA36343-1310) has been deposited withATCC on Mar. 30, 1998 and isassigned ATCC deposit no. 209718.

[1784] Analysis of the amino acid sequence of the full-length PRO324polypeptide suggests that it possesses significant sequence similarityto oxidoreductases, thereby indicating that PRO324 may be a noveloxidoreductase homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO324 amino acid sequence and the following Dayhoffsequences, B61209, A69965, YQJQ_BACSU, D69930, S76124, FABG_(—) ECOLI,C70023, S77280, FABG_VIBHA and MTV013_(—)6.

Example 22 Isolation of cDNA Clones Encoding Human PRO351

[1785] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA35950. Based on the DNA35950consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO351.

[1786] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-CCTGTGCTGTGCCTCGAGCCTGAC-3′ (SEQ ID NO:133) reverse PCR primer5′-GTGGGCAGCAGTTAGCACCGCCTC-3′ (SEQ ID NO:134)

[1787] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35950 sequence which had the followingnucleotide sequence

[1788] hybridization probe

[1789] 5′-GGCTGGCATCATCAGCTTTGCATCAAGCTGTGCCCAGGAGGACGC-3′ (SEQ IDNO:135)

[1790] In order ta screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplifilcation with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO351gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB230).

[1791] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO351 [herein designated as UNQ308(DNA40571-1315)] (SEQ ID NO:131) and the derived protein sequence forPRO351.

[1792] The entire nucleotide sequence of UNQ308 (DNA40571-1315) is shownin FIG. 48 (SEQ ID NO:131). Clone UNQ308 (DNA40571-1315) contains twoopen reading frames with an apparent translational initiation site atnucleotide positions 189-191 and a second open reading frame beginningat nucleotide 470, with the two open reading frames ending at the stopcodons at nucleotide positions 363-365 and 2009-2011, respectively (FIG.48). The predicted polypeptide precursor is 571 amino acids long (FIG.49). Important regions of the amino acid sequence of PRO351 include thesignal peptide, regions having sequence similarity to serine proteasesof the trypsin family, two N-glycosylation sites, and three Kringledomains. Clone UNQ308 (DNA40571-1315) has been deposited with ATCC andis assigned ATCC deposit no. 209784.

Example 23 Isolation of cDNA Clones Encoding Human PRO352

[1793] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA36950. Based on the DNA36950consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO352.

[1794] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 5′-CTGGCACAGCTCAACCTCATCTGG-3′ (SEQ ID NO:138) forward PCRprimer 2 5′-GCTGTCTGTCTGTCTCATTG-3′ (SEQ ID NO:139) forward PCR primer 35′-GGACACAGTATACTGACCAC-3′ (SEQ ID NO:140) reverse PCR primer 15′-TGCGAACCAGGCAGCTGTAAGTGC-3′ (SEQ ID NO:141) reverse PCR primer 25′-TGGAAGAAGAGGGTGGTGATGTGG-3′ (SEQ ID NO:142)

[1795] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA36950 sequence which had the followingnucleotide sequence

[1796] hybridization probe

[1797] 5′-CAGCTGACAGACACCAAACAGCTGGTGCACAGTTTCACCGAAGGC-3′ (SEQ IDNO:143)

[1798] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO352 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1799] DNA sequencing of the clones isolated as descnwed above gave thefull-length DNA sequence for PRO352 [herein designated as UNQ309(DNA41386-1316)1 (SEQ ID NO:136) and the derived protein sequence forPRO352.

[1800] The entire nucleotide sequence of UNQ309 (DNA4138&1316) is shownin FIG. 50 (SEQ ID NO:136). Clone UNQ309 (DNA41386-1316) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 152-154 and ending at the stop codon atnucleotide positions 1100-1102 (FIG. 50). The predicted polypeptideprecursor is 316 amino acids long (FIG. 51). The full-length PRO352protein shown in FIG. 2 has an estimated pI of about 4.62. Analysis ofthe full-length PRO352 sequence evidences the presence of a signalpeptide from about amino acid 1 to about amino acid 28, a transmembranedomain from about amino acid 251 to about amino acid 270, potentialN-glycosylation sites from about amino acid 91 to about amino acid 94,about amino acid 104 to about amino acid 107, about amino acid 189 toabout amino acid 192 and about amino acid 215 to about amino acid 218and a region having homology to immunoglobulins and MHC from about aminoacid 217 to about amino acid 234. Clone UNQ309 (DNA41386-1316) has beendeposited with ATCC on Mar. 26, 1998 and is assigned ATCC deposit no.209703.

[1801] Analysis of the amino acid sequence of the full-length PRO352polypeptide suggests that it possesses significant sequence similarityto the butyrophilin protein, thereby indicating that PRO352 is a novelbutyrophilin homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO352 amino acid sequence and the following Dayhoffsequences, BUTY_HUMAN, HSB73_(—)1, GGCD80_(—)1, I46690, A33_HUMAN,P_R67988, CD86_MOUSE, P_R71360, B39371 and D50558_(—)1.

Example 24 Isolation of cDNA Clones Encoding Human PRO381

[1802] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA39651. Based on the DNA39651consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO381.

[1803] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CTTTCCTTGCTTCAGCAACATGAGGC-3′ (SEQ ID NO:146)reverse PCR primer 5′-GCCCAGAGCAGGAGGAATGATGAGC-3′ (SEQ ID NO:147)

[1804] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA39651 sequence which had the followingnucleotide sequence

[1805] hybridization probe

[1806] G5′-GTGGAACGCGGTCTTGACTCTGTTCGTCACTTTGATTGGGGCTG-3′ (SEQ IDNO:148)

[1807] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO381 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1808] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO381 [herein designated as UNQ322(DNA44194-1317)] (SEQ ID NO:144) and the derived protein sequence forPRO381.

[1809] The entire nucleotide sequence of UNQ322 (DNA44194-1317) is shownin FIG. 52 (SEQ ID NO:144). Clone UNQ322 (DNA44194-1317) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 174-176 and ending at the stop codon atnucleotide positions 807-809 (FIG. 52). The predicted polypeptideprecursor is 211 amino acids long (FIG. 53). The full-length PRO381protein shown in FIG. 53 has an estimated molecular weight of about24,172 daltons and a pI of about 5.99. Analysis of the full-lengthPRO381 polypeptide shown in FIG. 53 (SEQ ID NO:145) evidences thepresence of the following: a signal peptide from about amino acid 1 toabout amino acid 20, a potential N-glycosylation site from about aminoacid 176 to about amino acid 179, potential casein kinase IIphosphorylation sites from about amino acid 143 to about amino acid 146,from about amino acid 156 to about amino acid 159, from about amino acid178 to about amino acid 181, and from about amino acid 200 to aboutamino acid 203, an endoplasmic reticulum targeting sequence from aboutamino acid 208 to about amino acid 211, FKBP-type peptidyl-prolylcis-trans isomerase sites from about amino acid 78 to about amino acid114 and from about amino acid 118 to about amino acid 131, EF-handcalcium binding domains from about amino acid 191 to about amino acid203, from about amino acid 184 to about amino acid 203 and from aboutamino acid 140 to about amino acid 159, and an S-100/ICaBP type calciumbinding domain from about amino acid 183 to about amino acid 203. CloneUNQ322 (DNA44194-1317) has been deposited with ATCC on Apr. 28, 1998 andis assigned ATCC deposit no. 209808.

[1810] Analysis of the amino acid sequence of the full-length PRO381polypeptide suggests that it possesses significant sequence similarityto FKBP immunophilin proteins, thereby indicating that PRO381 may be anovel FKBP immunophilin homolog. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO381 amino acid sequence and the followingDayhoff sequences, AF040252_(—)1, 149669, P_R93551, S71238,CELC05C8_(—)1, CEU27353_(—)1, MIP_TRYCR, CEZC455_(—)3, FKB4HUMAN andI40718.

Example 25 Isolation of cDNA Clones Encoding Human PRO386

[1811] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA40674. Two proprietaryGenentech EST sequences were employed in the consensus sequenceassembly, wherein those EST sequences are herein designated DNA23350(FIG. 56; SEQ ID NO:151) and DNA23536 (FIG. 57; SEQ ID NO:152). Based onthe DNA40674 consensus sequence, oligonucleotides were synthesized: 1)to identify by PCR a cDNA library that contained the sequence ofinterest, and 2) for use as probes to isolate a clone of the full-lengthcoding sequence for PRO386.

[1812] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-ACGGAGCATGGAGGTCCACAGTAC-3′ (SEQ ID NO:153)reverse PCR primer 5′-GCACGTTTCTCAGCATCACCGAC-3′ (SEQ ID NO:154)

[1813] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA40674 sequence which had the followingnucleotide sequence

[1814] hybridization probe

[1815] 5′-CGCCTGCCCTGCACCTTCAACTCCTGCTACACAGTGAACCACAAACAGTT-3′ (SEQ IDNO:155)

[1816] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO386 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal braintissue (LIB153).

[1817] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO386 herein designated as UNQ326(DNA45415-1318)] (SEQ ID NO:149) and the derived protein sequence forPRO386.

[1818] The entire nucleotide sequence of UNQ326 (DNA45415-1318) is shownin FIG. 54 (SEQ ID NO:149). Clone UNQ326 (DNA45415-1318) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 146-148 and ending at the stop codon atnucleotide positions 791-793 (FIG. 54). The predicted polypeptideprecursor is 215 amino acids long (FIG. 55). The full-length PRO386protein shown in FIG. 55 has an estimated molecular weight of about24,326 daltons and a pI of about 6.32. Analysis of the full-lengthPRO386 sequence shown in FIG. 55 (SEQ ID NO:150) evidences the presenceof the following: a signal peptide from about amino acid 1 to aboutamino acid 20, a transmembrane domain from about amino acid 161 to aboutamino acid 179, an immunoglobulin-like fold from about amino acid 83 toabout amino acid 127 and potential N-glycosylation sites from aboutamino acid 42 to about amino acid 45, from about amino acid 66 to aboutamino acid 69 and from about amino acid 74 to about amino acid 77. CloneUNQ326 (DNA45415-1318) has been deposited with ATCC on Apr. 28, 1998 andis assigned ATCC deposit no. 209810.

[1819] Analysis of the amino acid sequence of the full-length PRO386polypeptide suggests that it possesses significant sequence similarityto the sodium channel beta-2 subunit, thereby indicating that PRO386 isa novel homolog thereof. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO386 amino acid sequence and the following Dayhoffsequences, A57843, MYP0_HUMAN, GEN14531, JC4024, HS46KDA_(—)1,HSU90716_(—)1, D86996_(—)2, MUSIGLVD_(—)1, DMU42769_(—)1 and S19247.

Example 26 Isolation of cDNA Clones Encoding Human PRO540

[1820] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA39631. Based on the DNA39631consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO540.

[1821] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-CTGGGGCTACACACGGGGTGAGG-3′ (SEQ ID NO:158) reverse PCR primer5′-GGTGCCGCTGCAGAAAGTAGAGCG-3′ (SEQ ID NO:159)

[1822] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA40654 sequence which had the followingnucleotide sequence

[1823] hybridization probe

[1824] 5′-GCCCCAAATGAAAACGGGCCCTACTTCCTGGCCCTCCGCGAGATG-3′ (SEQ IDNO:160)

[1825] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO540gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1826] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO540 [herein designated as UNQ341(DNA44189-1322)] (SEQ ID NO:156) and the derived protein sequence forPRO540.

[1827] The entire nucleotide sequence of UNQ341 (DNA44189-1322) is shownin FIG. 58 (SEQ ID NO:156). Clone UNQ341 (DNA44189-1322) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 21-23 and ending at the stop codon at nucleotidepositions 1257-1259 (FIG. 58). The predicted polypeptide precursor is412 amino acids long (FIG. 59). The full-length PRO540 protein shown inFIG. 59 has an estimated molecular weight of about 46,658 daltons and apI of about 6.65. Important regions of the amino acid sequence of PRO540include the signal peptide, potential N-glycosylation sites, a potentiallipid substrate binding site, a sequence typical of lipases and serineproteins, and a beta-transducin family Trp-Asp repeat. Clone UNQ341(DNA44189-1322) has been deposited with ATCC and is assigned ATCCdeposit no. 209699.

Example 27 Isolation of cDNA Clones Encoding Human PRO615

[1828] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA42240. Based on the DNA42240consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO615.

[1829] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-TGGTCTTCGCCTTGATCGTGTTCT-3′ (SEQ ID NO:163) forward PCR primer5′-GTGTACTGAGCGGCGGTTAG-3′ (SEQ ID NO:164) reverse PCR primer5′-CTGAAGGTGATGGCTGCCCTCAC-3′ (SEQ ID NO:165) reverse PCR primer5′-CCAGGAGGCTCATGGGAAAGTCC-3′ (SEQ ID NO:166)

[1830] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA42240 sequence which had the followingnucleotide sequence:

[1831] hybridization probe

[1832] 5′-CCACGAGTCTAAGCAGATGTACTGCGTGTTCAACCGCAACGAGGATGCCT-3′ (SEQ IDNO:167)

[1833] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO615gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrowtissue (LIB255).

[1834] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO615 [herein designated as UNQ352(DNA43304-1323)] (SEQ ID NO:161) and the derived protein sequence forPRO615.

[1835] The entire nucleotide sequence of UNQ352 (DNA48304-1323) is shownin FIG. 60 (SEQ ID NO:161). Clone UNQ352 (DNA48304-1323) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 51-53 and ending at the stop codon at nucleotidepositions 723-725 (FIG. 60). The predicted polypeptide precursor is 224amino acids long (FIG. 61). The full-length PRO615 protein shown in FIG.61 has an estimated molecular weight of about 24,810 daltons and a pI ofabout 4.75. Important regions of the amino acid sequence of PRO615include a type II transmembrane domain, corresponding to about aminoacids 24-43, other transmembrane domains, corresponding to about aminoacids 74-90, 108-126, and 145-161, respectively, and a potentialN-glycosylation site, corresponding to about amino acids 97-100. CloneUNQ352 (DNA48304-1323) has been deposited with ATCC and is assigned ATCCdeposit no. 209811.

Example 28 Isolation of cDNA Clones Encoding Human PRO618

[1836] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA30900. Based on the DNA30900consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO618.

[1837] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-TAACAGCTGCCCACTGCTTCCAGG-3′ (SEQ ID NO:171) reverse PCR primer5′-TAATCCAGCAGTGCAGGCCGGG-3′ (SEQ ID NO:172)

[1838] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA30900 sequence which had the followingnucleotide sequence

[1839] hybridization probe

[1840] 5′-ATGGCCTCCACGGTGCTGTGGACCGTGTTCCTGGGCAAGGTGTGGCAGAA-3′ (SEQ IDNO:173)

[1841] Screening of the above described library gave rise to the partialcDNA clone designated herein DNA35597 (SEQ ID NO:170). Extension of thissequence using repeated cycles of BLAST and phrap gave rise to anucleotide sequence designated herein as DNA43335. Primers based uponthe DNA43335 consensus sequence were then prepared as follows. forwardPCR primer 5′-TGCCTATGCACTGAGGAGGCAGAAG-3′ (SEQ ID NO:174) reverse PCRprimer 5′-AGGCAGGGACACAGAGTCCATTCAC-3′ (SEQ ID NO:175)

[1842] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA43335 sequence which had the followingnucleotide sequence

[1843] hybridization probe

[1844] 5′-AGTATGATTTGCCGTGCACCCAGGGCCAGTGGACGATCCAGAACAGGAGG-3′ (SEQ IDNO:176)

[1845] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate full length clones encodingthe PRO618 gene using the second probe oligonucleotide and one of thesecond set of PCR primers. RNA for construction of the cDNA librarieswas isolated from human fetal liver tissue (LIB229).

[1846] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO618 herein designated as UNQ354(DNA49152-1324)] (SEQ ID NO:168) and the derived protein sequence forPRO618.

[1847] The entire nucleotide sequence of UNQ354 (DNA49152-1324) is shownin FIG. 62 (SEQ ID NO:168). Clone UNQ354 (DNA49152-1324) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 73-75 and ending at the stop codon at nucleotidepositions 2479-2481 (FIG. 62). The predicted polypeptide precursor is802 amino acids long (FIG. 63). The full-length PRO618 protein shown inFIG. 63 has an estimated molecular weight of about 88,846 daltons and apI of about 6.41. Important regions of the amino acid sequence of PRO618include type II transmembrane domain, a sequence typical of a protease,trypsin family, histidine active site, multiple N-glycosylation sites,two sequences typical of a Kringle domain, two regions having sequencesimilarity to Kallikrein light chain, and a region having sequencesimilarity to low-density lipoprotein receptor. Clone UNQ354(DNA49152-1324) has been deposited with ATCC and is assigned ATCCdeposit no. 209813.

Example 29 Isolation of cDNA Clones Encoding Human PRO719

[1848] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA44851. Based on the DNA44851consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO719.

[1849] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-GTGAGCATGAGCGAGCCGTCCAC-3′ (SEQ ID NO:179) reversePCR primer 5′-GCTATTACAACGGTTCTTGCGGCAGC-3′ (SEQ ID NO:180)

[1850] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA44851 sequence which had the followingnucleotide sequence

[1851] hybridization probe

[1852] 5′-TTGACTCTCTGGTGAATCAGGACAAGCCGAGTTTTGCCTTCCAG-3′ (SEQ IDNO:181)

[1853] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO719 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human placentatissue (LIB90).

[1854] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO719 [herein designated as UNQ387(DNA49646-1327)] (SEQ ID NO:177) and the derived protein sequence forPRO719.

[1855] The entire nucleotide sequence of UNQ387 (DNA49646-1327) is shownin FIG. 65 (SEQ ID NO:177). Clone UNQ387 (DNA49646-1327) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 223-225 and ending at the stop codon atnucleotide positions 1285-1287 (FIG. 65). The predicted polypeptideprecursor is 354 amino acids long (FIG. 66). The full-length PRO719protein shown in FIG. 66 has an estimated molecular weight of about39,362 daltons and a pI of about 8.35. Analysis of the full lengthPRO719 sequence evidences the presence of a signal peptide from aboutamino acid 1 to about amino acid 16 as shown in FIG. 66 (SEQ ID NO:178),a lipase-associated serine-containing active site at about amino acid163 to about amino acid 172, and two potential N-glycosylation sitesfrom about amino acid 80 to about amino acid 83 and about amino acid 136to about amino acid 139. Clone UNQ387 (DNA49646-1327) has been depositedwith ATCC on Mar. 26, 1998 and is assigned ATCC deposit no. 209705.

[1856] Analysis of the amino acid sequence of the full-length PRO719polypeptide suggests that it possesses significant sequence similarityto the lipoprotein lipase Hprotein, thereby indicating that PRO719 maybe a novel lipoprotein lipase homolog. More specifically, an analysis ofthe Dayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO719 amino acid sequence and the followingDayhoff sequences, LIPL_HUMAN, LIPH_HUMAN, D83548_(—)1, A24059_(—)1,PR30740, D88666_(—)1, A43357, A46696, B43357 and A49488.

Example 30 Isolation of cDNA Clones Encoding Human PRO724

[1857] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA35603. Based on the DNA35603consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO724.

[1858] Pairs of PCR primers (forward and reverse) were synthesized:forward PCR primer 1 5′-GGCTGTCACTGTGGAGACAC-3′ (SEQ ID NO:184) forwardPCR primer 2 5′-GCAAGGTCATTACAGCTG-3′ (SEQ ID NO:185) reverse PCR primer1 5′-AGAACATAGGAGCAGTCCCACTC-3′ (SEQ ID NO:186) reverse PCR primer 25′-TGCCTGCTGCTGCACAATCTCAG-3′ (SEQ ID NO:187)

[1859] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35603 sequence which had the followingnucleotide sequence

[1860] hybridization probe

[1861] 5′-GGCTATTGCTTGCCTTGGGACAGACCCTGTGGCTTAGGCTCTGGC-3′ (SEQ IDNO:188)

[1862] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pairs identified above. A positivelibrary was then used to isolate clones encoding the PRO724 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal lungtissue (LIB26).

[1863] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO724 [herein designated as UNQ389(DNA49631-1328)] (SEQ ID NO:182) and the derived protein sequence forPRO724.

[1864] The entire nucleotide sequence of UNQ389 (DNA49631-1328) is shownin FIG. 67 (SEQ ID NO:182). Clone UNQ389 (DNA49631-1328) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 546-548 and ending at the stop codon atnucleotide positions 2685-2687 (FIG. 67). The predicted polypeptideprecursor is 713 amino acids long (FIG. 68). The full-length PRO724protein shown in FIG. 68 has an estimated molecular weight of about76,193 daltons and a pI of about 5.42. Analysis of the full-lengthPRO724 amino acid sequence shown in FIG. 68 (SEQ ID NO:183) evidencesthe presence of the following: a signal peptide from about amino acid 1to about amino acid 16, a transmembrane domain from about amino acid 442to about amino acid 462 and LDL receptor class A domain regions fromabout amino acid 152 to about amino acid 171, about amino acid 331 toabout amino acid 350, about amino acid 374 to about amino acid 393 andabout amino acid 411 to about amino acid 430. Clone UNQ389(DNA49631-1328) has been deposited with ATCC on Apr. 28, 1998 and isassigned ATCC deposit no. 209806 Analysis of the amino acid sequence ofthe full-length PRO724 polypeptide suggests that it possessessignificant sequence similarity to the human LDL receptor protein,thereby indicating that PRO724 may be a novel LDL receptor homolog. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant homology between the PRO724 aminoacid sequence and the following Dayhoff sequences, P_R48547,MMAM2R_(—)1, LRP2_RAT, P_R60517, P_R47861, P_R05533, A44513_(—)1,A30363, P_R74692 and LMLIPOPHO_(—)1.

Example 31 Isolation of cDNA Clones Encoding Human PRO772

[1865] One cDNA sequence was isolated in the amylase screen described inExample 2, wherein that cDNA sequence is herein designated DNA43509 (seeFIG. 71). Based on the DNA43509 sequence, oligonucleotide probes weregenerated and used to screen a human fetal lung library (LIB25) preparedas described in paragraph 1 of Example 2 above. The cloning vector waspRK5B (pRK5B is a precursor of pRK5D that does not contain the SfiIsite; see, Holmes et al., Science, 253:1278-1280 (1991)), and the cDNAsize cut was less than 2800 bp.

[1866] A pair of PCR primers (forward and reverse) were synthesizedbased on the DNA43509 sequence: forward PCR primer5′-CGTTTTGCAGAACCTACTCAGGCAG-3′ (SEQ ID NO:192) reverse PCR primer5′-CCTCCACCAACTGTCAATGTTGTGG-3′ (SEQ ID NO:193)

[1867] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA43509 sequence which had the followingnucleotide sequence

[1868] hybridization probe

[1869] 5′-AAAGTGCTGCTGCTGGGTCTGCAGACGCGATGGATAACGT-3′ (SEQ ID NO:194)

[1870] Using the above described primers and library, a full lengthclone was identified that contained a single open reading frame with anapparent translational initiation site at nucleotide positions 131-133and ending at the stop codon found at nucleotide positions 587-589 (FIG.69; SEQ ID NO:189). The predicted polypeptide precursor is 152 aminoacids long, has a calculated molecular weight of approximnately 17,170daltons and an estimated pI of approximately 9.62. Analysis of thefull-length PRO772 sequence shown in FIG. 70 (SEQ ]ID NO:190) evidencesthe presence of the following: a potential type II transmembrane domainfrom about amino acid 26 to about amino acid 42, other potentialtransmembrane domains from about amino acid 44 to about amino acid 65,from about amino acid 81 to about amino acid 101 and from about aminoacid 109 to about amino acid 129, leucine zipper pattern sequences fromabout amino acid 78 to about amino acid 99 and from about amino 7 acid85 to about amino acid 106. Clone UNQ410 (DNA49645-1347) has beendeposited with ATCC on Apr. 28, 1998 and is assigned ATCC deposit no.209809.

[1871] Analysis of the amino acid sequence of the full-length PRO772polypeptide suggests that it possesses significant sequence similarityto the human A4 protein, thereby indicating that PRO772 may be a novelA4 protein homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO772 amino acid sequence and the following Dayhoffsequences, HSU93305_(—)1, A4P_HUMAN, CELB0454_(—)2, VPU_JSRV,CELC12D12_(—)2, OCCM_AGRT1, LBPHIG1E_(—)50, YIGK_(—) ECOLI, S76245 andP_R50807.

Example 32 Isolation of cDNA Clones Encoding Human PRO852

[1872] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA34364. Based on the DNA34364consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO852.

[1873] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 5′-CGCAGAAGCTACAGATTCTCG-3′ (SEQ ID NO:197) forward PCR primer2 5′-GGAAATTGGAGGCCAAAGC-3′ (SEQ ID NO:198) forward PCR Primer 35′-GGATGTAGCCAGCAACTGTG-3′ (SEQ ID NO:199) forward PCR primer 45′-GCCTTGGCTCGTTCTCTTC-3′ (SEQ ID NO:200) forward PCR primer 55′-GGTCCTGTGCCTGGATGG-3′ (SEQ ID NO:201) reverse PCR primer 15′-GACAAGACTACCTCCGTTGGTC-3′ (SEQ ID NO:202) reverse PCR primer 25′-TGATGCACAGTTCAGCACCTGTTG-3′ (SEQ ID NO:203)

[1874] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA34364 sequence which had the followingnucleotide sequence

[1875] hybridization probe

[1876] 5′-CGCTCCAAGGGCTrTGACGTCACAGTGAAGTACACACAAGGAAGCTG-3′ (SEQ IDNO:204)

[1877] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO852 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB228).

[1878] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO852 [herein designated as UNQ418(DNA45493-1349)] (SEQ ID NO:195) and the derived protein sequence forPRO852.

[1879] The entire nucleotide sequence of UNQ418 (DNA45493-1349) is shownin FIG. 72 (SEQ ID NO:195). Clone UNQ418 (DNA45493-1349) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 94-96 and ending at the stop codon at nucleotidepositions 16748-1650 (FIG. 72). The predicted polypeptide precursor is518 amino acids long (FIG. 73). The full-length PRO852 protein shown inFIG. 73 has an estimated molecular weight of about 56,180 daltons and apI of about 5.08. Analysis of the full-length PRO852 sequence shown inFIG. 73 (SEQ ID NO:196) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 20, atransmembrane domain from about amino acid 466 to about amino acid 494,potential N-glycosylation sites from about amino acid 170 to about aminoacid 173 and about amino acid 366 to about amino acid 369, leucinezipper sequence pattern blocks from about amino acid 10 to about aminoacid 31 and from about amino acid 197 to about amino acid 218 and blocksof amino acids having sequence homology to eukaryotic and viral aspartylproteases from about amino acid 109 to about amino acid 118, from aboutamino acid 252 to about amino acid 261 and from about amino acid 298 toabout amino acid 310. Clone UNQ418 (DNA45493-1349) has been depositedwith ATCC on Apr. 28, 1998 and is assigned ATCC deposit no. 209805.

[1880] Analysis of the amino acid sequence of the full-length PRO852polypeptide suggests that it possesses significant sequence similarityto various protease proteins, thereby indicating that PRO852 may be anovel protease protein or homolog thereof. More specifically, ananalysis of the Dayhoff database (version 35.45 SwissProt 35) evidencedsignificant homology between the PRO852 amino acid sequence and thefollowing Dayhoff sequences, PEPC_HUMAN, S66516, S66517, PEPE_CHICK,CATD^(—)HUMAN, P_R74207, CARP_YEAST, PEP2_RABIT, CATE_HUMAN andRENI_MOUSE.

Example 33 Isolation of cDNA Clones Encoding Human PRO853

[1881] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43050. Based on the DNA43050consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO853.

[1882] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-CTTCATGGCCTTGGACTTGGCCAG-3′ (SEQ ID NO:207) reverse PCR primer5′-ACGCCAGTGGCCTCAAGCTGGTTG-3′ (SEQ ID NO:208)

[1883] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA43050 sequence which had the followingnucleotide sequence

[1884] hybridization probe

[1885] 5′-CTTTCTGAGCTCTGAGCCACGGTTGGACATCCTCATCCACAATGC-3′ (SEQ IDNO:209)

[1886] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO853gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB228).

[1887] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO853 [herein designated as UNQ419(DNA48227-1350)] (SEQ 1D NO:205) and the derived protein sequence forPRO853.

[1888] The entire nucleotide sequence of UNQ419 (DNA48227-1350) is shownin FIG. 74 (SEQ ID NO:205). Clone UNQ419 (DNA48227-1350) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 128-130 and ending at the stop codon atnucleotide positions 1259-1261 (FIG. 74). The predicted polypeptideprecursor is 377 amino acids long (FIG. 75). The full-length PRO853protein shown in FIG. 75 has an estimated molecular weight of about40,849 daltons and a pi of about 7.98. Important regions of the aminoacid sequence of PRO853 include the signal peptide, corresponding toamino acids from about 1 to about 16 of SEQ ID NO:206, theglycosaminoglycan attachment site, corresponding to amino acids fromabout 46 to about 49 of SEQ ID NO:206, and two sequences typical of theshort-chain alcohol dehydrogenase family, corresponding to amino acidsfrom about 37 to about 49 and about 114 to about 124 of SEQ ID NO:206,respectively. Clone UNQ419 (DNA48227-1350) has been deposited with ATCCand is assigned ATCC deposit no. 209812.

Example 34 Isolation of cDNA Clones Encoding Human PRO860

[1889] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA38137. Based on the DNA38137consensu sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO860.

[1890] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-GAAGGGACCTACATGTGTGTGGCC-3′ (SEQ ID NO:212) reverse PCR primer5′-ACTGACCTTCCAGCTGAGCCACAC-3′ (SEQ ID NO:213)

[1891] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA40654 sequence which had the followingnucleotide sequence

[1892] hybridization probe

[1893] 5′-AGGACTACACGGAGCCTGTGGAGCTTCTGGCTGTGCGAATTCAGCTGGAA-3′ (SEQ IDNO:214)

[1894] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO860gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal lungtissue (LIB26).

[1895] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO860 [herein designated as UNQ421(DNA41404-1352)] (SEQ ID NO:210) and the derived protein sequence forPRO860.

[1896] The entire nucleotide sequence of UNQ421 (DNA41404-1352) is shownin FIG. 76 (SEQ ID NO:210). Clone UNQ421 (DNA41404-1352) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 58-60 and ending at the stop codon at nucleotidepositions 3013-3015 (FIG. 76). The predicted polypeptide precursor is985 amino acids long (FIG. 77). The full-length PRO860 protein shown inFIG. 77 has an estimated molecular weight of about 105,336 daltons and apI of about 6.55. Important regions of the amino acid sequence of PRO860include the transmembrane region corresponding to about amino acids448-467, the extracellular domain, corresponding to amino acids about1-447, several N-glycosylation sites, numerous N-myristoylation sitesand a sequence typical of phosphotyrosine interaction domain proteins .Clone UNQ421 (DNA41404-1352) has been deposited with ATCC and isassigned ATCC deposit no. 209844.

Example 35 Isolation of cDNA Clones Encoding Human PRO846

[1897] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA39949. Based on the DNA39949consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO846.

[1898] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-CCCTGCAGTGCACCTACAGGGAAG-3′ (SEQ ID NO:217) reverse PCR primer5′-CTGTCTTCCCCTGCTTGGCTGTGG-3′ (SEQ ID NO:218)

[1899] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA39949 sequence which had the followingnucleotide sequence

[1900] hybridization probe

[1901] 5′-GGTGCAGGAAGGGTGGGATCCTCTTCTCTCGCTGCTCTGGCCACATC-3′ (SEQ IDNO:219)

[1902] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO546gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1903] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO846 [herein designated as UNQ422(DNA44196-1353)] (SEQ ID NO:215) and the derived protein sequence forPRO846.

[1904] The entire nucleotide sequence of UNQ422 (DNA44196-1353) is shownin FIG. 78 (SEQ ID NO:215). Clone UNQ422 (DNA44196-1353) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 25-27 and ending at the stop codon at nucleotidepositions 1021-1023 (FIG. 78). The predicted polypeptide precursor is332 amino acids long (FIG. 79). The full-length PRO846 protein shown inFIG. 79 has an estimated molecular weight of about 36,143 daltons and apI of about 5.89. Important regions of the amino acid sequence of PRO846include the signal peptide, the transmembrane domain, an N-glycosylationsite, a sequence typical of fibrinogen beta and gamma chains C-terminaldomain, and a sequence typical of Ig like V-type domain as shown in FIG.79. Clone UNQ422 (DNA44196-1353) has been deposited with ATCC and isassigned ATCC deposit no. 209847.

Example 36 Isolation of cDNA Clones Encoding Human PRO862

[1905] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA47370. Based on the DNA47370consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO862.

[1906] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′GGGATCATGTTGTTGGCCCTGGTC-3′ (SEQ ID NO:222) reverse PCR primer5′-GCAAGGCAGACCCAGTCAGCCAG-3′ (SEQ ID NO:223)

[1907] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA47370 sequence which had the followingnucleotide sequence

[1908] hybridization probe

[1909] 5′-CTGCCTGCTACCCTCCAAGTGAGGCCAAGCTCTACGGTCGTTGTG-3′ (SEQ IDNO:225)

[1910] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO862gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human pancreastissue (LIB55).

[1911] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO862 [herein designated as UNQ424(DNA52187-1354)] (SEQ ID NO:220) and the derived protein sequence forPRO862.

[1912] The entire nucleotide sequence of UNQ424 (DNA52187-1354) is shownin FIG. 80 (SEQ ID NO:220). Clone UNQ424 (DNA52187-1354) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 410-412 and ending at the stop codon atnucleotide positions 848-850 (FIG. 80). The predicted polypeptideprecursor is 146 amino acids long (FIG. 81). The full-length PRO862protein shown in FIG. 81 has an estimated molecular weight of about16,430 daltons and a pI of about 5.05. Important regions of the aminoacid sequence of PRO862 include the signal peptide, an N-myristoylationsite, and sequences having similarity to region toAlpha-lactalbumin/lysozyme C proteins as shown in FIG. 81. Clone UNQ424(DNA52187-1354) has been deposited with the ATCC and is assigned ATCCdeposit no. 209845.

Example 37 Isolation of cDNA Clones Encoding Human PRO864

[1913] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA40666. Based on the DNA40666consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO864.

[1914] Forward and reverse PCR primers were synthesized: forward PCRprimer 5′-GCTGCAGCTGCAAATTCCACTGG-3′ (SEQ ID NO:227) reverse PCR primer5′-TGGTGGGAGACTGTTTAAATTATCGGCC-3′ (SEQ ID NO:228)

[1915] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA40666 sequence which had the followingnucleotide sequence

[1916] hybridization probe

[1917] 5′-TGCTTCGTCAAGTGCCGGCAGTGCCAGCGGCrCGTGGAGTT-3′ (SEQ ID NO:229)

[1918] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO864gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal braintissue (LIB153).

[1919] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO864 [herein designated as UNQ426(DNA48328-1355)] (SEQ ID NO:225) and the derived protein sequence forPRO864.

[1920] The entire nucleotide sequence of UNQ426 (DNA48328-1355) is shownin FIG. 82 (SEQ ID NO:225). Clone UNQ426 (DNA48328-1355) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 37-39 and ending at the stop codon at nucleotidepositions 1090-1092 (FIG. 82). The predicted polypeptide precursor is351 amino acids long (FIG. 83). The full-length PRO864 protein shown inFIG. 83 has an estimated molecular weight of about 39,052 and apI ofabout 8.97. Important regions of the amino acid sequence of PRO864include the signal peptide, two N-glycosylation sites, a Wnt-1 familysignature sequence, and sequence regions homologous to Wnt-1 familyproteins as shown in FIG. 83. Clone UNQ426 (DNA48328-1355) has beendeposited with ATCC and is assigned ATCC deposit no. 209843.

Example 38 Isolation of cDNA Clones Encoding Human PRO792

[1921] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA38106. Based on the DNA38106consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO792.

[1922] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-GCGAGAACTGTGTCATGATGCTGC-3′ (SEQ ID NO:232)reverse PCR primer 5′-GTTTCTGAGACTCAGCAGCGGTGG-3′ (SEQ ID NO:233)

[1923] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA38106 sequence which had the followingnucleotide sequence

[1924] hybridization probe

[1925] 5′-CACCGTGTGACAGCGAGAAGGACGGCTGGATCTGTGAGAAAAGGCACAAC-3′ (SEQ IDNO:234)

[1926] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO792 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrowtissue (LIB255).

[1927] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO792 [herein designated as UNQ431(DNA56352-1358)] (SEQ ID NO:230) and the derived protein sequence forPRO792.

[1928] The entire nucleotide sequence of UNQ431 (DNA56352-1358) is shownin FIG. 84 (SEQ ID NO:230). Clone UNQ431 (DNA56352-1358) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 67-69 and ending at the stop codon at nucleotidepositions 946-948 (FIG. 84). The predicted polypeptide precursor is 293amino acids long (FIG. 85). The full-length PRO792 protein shown in FIG.85 has an estimated molecular weight of about 32,562 daltons and a pI ofabout 6.53. Analysis of the full-length PRO792 sequence shown in FIG. 85(SEQ ID NO:231) evidences the presence of the following: a type IItransmembrane domain from about amino acid 31 to about amino acid 54,potential N-glycosylation sites from about amino acid 73 to about aminoacid 76 and from about amino acid 159 to about amino acid 162, a leucinezipper amino acid sequence pattern from about amino acid 102 to aboutamino acid 123, potential N-myristolation sites from about amino acid 18to about amino acid 23, from about amino acid 133 to about amino acid138 and from about amino acid 242 to about amino acid 247 and a C-typelectin domain signature block from about amino acid 264 to about aminoacid 287. Clone UNQ431 (DNA56352-1358) has been deposited with ATCC onMay 6, 1998 and is assigned ATCC deposit no. 209846.

[1929] Analysis of the amino acid sequence of the full-length PRO792polypeptide suggests that it possesses significant sequence similarityto the CD23 protein, thereby indicating that PRO792 may be a novel CD23homolog. More specifically, an analysis of the Dayhoff database (version35.45 SwissProt 35) evidenced significant homology between the PRO792amino acid sequence and the following Dayhoff sequences, S34198,A07100_(—)1, A05303_(—)1, P_R41689, P_P82839, A10871_(—)1, P_R12796,P_R47199, A46274 and P_R32188.

Example 39 Isolation of cDNA Clones Encoding Human PRO866

[1930] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA44708. Based on the DNA44708consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO866.

[1931] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 5′-CAGCACTGCCAGGGGAAGAGGG-3′ (SEQ ID NO:237) forward PCR primer2 5′-CAGGACTCGCTACGTCCG-3′ (SEQ ID NO:238) forward PCR primer 35′-CAGCCCCTTCTCCTCCTTTCTCCC-3′ (SEQ ID NO:239) reverse PCR primer 15′-GCAGTTATCAGGGACGCACTCAGCC-3′ (SEQ ID NO:240) reverse PCR primer 25′-CCAGCGAGAGGCAGATAG-3′ (SEQ ID NO:241) reverse PCR primer 35′-CGGTCACCGTGTCCTGCGGGATG-3′ (SEQ ID NO:242)

[1932] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA44708 sequence which had the followingnucleotide sequence

[1933] hybridization probe

[1934] 5′-CAGCCCCTTCTCCTCCTTTCTCCCACGTCCTATCTGCCTCTC-3′ (SEQ ID NO:243)

[1935] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO866gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB228).

[1936] DNA sequencing of the clones isolated as described above gave thefiWllIength DNA sequence for PRO866 [herein designated as UNQ435(DNA53971-1359)] (SEQ ID NO:235) and the derived protein sequence forPRO866.

[1937] The entire nucleotide sequence of UNQ435 (DNA53971-1359) is shownin FIG. 86 (SEQ ID NO:235). Clone UNQ435 (DNA53971-1359) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 275-277 and ending at the stop codon atnucleotide positions 1268-1270 (FIG. 86). The predicted polypeptideprecursor is 331 amino acids long (FIG. 87). The full-length PRO866protein shown in FIG. 87 has an estimated molecular weight of about35,844 daltons and a pI of about 5.45. Analysis of the full-lengthPRO866 sequence shown in FIG. 87 (SEQ ID NO:236) evidences the presenceof the following: a signal peptide from about amino acid 1 to aboutamino acid 26. Clone UNQ435 (DNA53971-1359) has been deposited with ATCCon Apr. 7, 1998 and is assigned ATCC deposit no. 209750.

[1938] Analysis of the amino acid sequence of the full-length PRO866polypeptide suggests that it possesses significant sequence similarityto the mindin/spondin family of proteins, thereby indicating that PRO866may be a novel mindin homolog. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO866 amino acid sequence and the followingDayhoff sequences, AB006085_(—)1, AB006084_(—)1, AB006086_(—)1,AF017267_(—)1, CWU42213_(—)1, AC004160_(—)1, CPMICRP1, S49108, A48569and I46687.

Example 40 Isolation of cDNA Clones Encoding Human PRO871

[1939] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA40324. Based on the DNA40324consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO871.

[1940] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 5′-TGCGGAGATCCTACTGGCACAGGG-3′ (SEQ ID NO:246) forward PCRprimer 2 5′-CGAGTTAGTCAGAGCATG-3′ (SEQ ID NO:247) forward PCR primer 35′-CAGATGGTGCTGTTGCCG-3′ (SEQ ID NO:248) reverse PCR primer 15′-CAACTGGAACAGGAACTGAGATGTGGATC-3′ (SEQ ID NO:249) reverse PCR primer 25′-CTGGTTCAGCAGTGCAAGGGTCTG-3′ (SEQ ID NO:250) reverse PCR primer 35′-CCTCTCCGATTAAAACGC-3′ (SEQ ID NO:251)

[1941] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA40324 sequence which had the followingnucleotide sequence

[1942] hybridization probe

[1943] 5′-GAGAGGACTGGTTGCCATGGCAAATGCTGGTTCTCATGATAATGG-3′ (SEQ IDNO:252)

[1944] In order to screen several libraries for a source of afiul-length clone, DNA from the libraries was screened by PCRamplification with one of the PCR primer pairs identified above. Apositive library was then used to isolate clones encoding the PRO871gene using the probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1945] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO871 therein designated as UNQ438(DNA50919-1361)] (SEQ ID NO:244) and the derived protein sequence forPRO871.

[1946] The entire nucleotide sequence of UNQ438 (DNA50919-1361) is shownin FIG. 88 (SEQ ID NO:244). Clone UNQ438 (DNA50919-1361) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 191-193 and ending at the stop codon atnucleotide positions 1607-1609 (FIG. 88). The predicted polypeptideprecursor is 472 amino acids long (FIG. 89). The full-length PRO871protein shown in FIG. 89 has an estimated molecular weight of about53,847 daltons and a pI of about 5.75. Analysis of the full-lengthPRO871 sequence shown in FIG. 89 (SEQ ID NO:245) evidences the presenceof the following: a signal peptide from about amino acid 1 to aboutamino acid 21, potential N-glycosylation sites from about amino acid 109to about amino acid 112 and from about amino acid 201 to about aminoacid 204, a cyclophilin-type peptidy-prolyl cis-trans isomerasesignature sequence from about amino acid 49 to about amino acid 66 andregions that are homologous to cyclophilin-type peptidy-prolyl cis-transisomerases from about amino acid 96 to about amino acid 140, from aboutamino acid 49 to about amino acid 89 and from about amino acid 22 toabout amino acid 51. Clone UNQ438 (DNA50919-1361) has been depositedwith ATCC on May 6, 1998 and is assigned ATCC deposit no. 209848.

[1947] Analysis of the amino acid sequence of the full-length PRO871polypeptide suggests that it possesses significant sequence similarityto the cyclophilin family of proteins, thereby indicating that PRO871may be a novel cyclophilin protein family member. More specifically, ananalysis of the Dayhoff database (version 35.45 SwissProt 35) evidencedsignificant homology between the PRO871 amino acid sequence and thefollowing Dayhoff sequences, SPBC16H5_(—)5, S64705, YAL5_SCHPO,CYP4_CAEEL, CELC34D4_(—)7, CYPA_CAEEL, HUMORF006_(—)1, CYPI_MYCTU,AF043642_(—)1 and HSSRCYP_(—)1.

Example 41 Isolation of cDNA Clones Encoding Human PRO873

[1948] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA39621. Based on the DNA39621consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO873.

[1949] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-AGGTGCCTGCAGGAGTCCTGGGG-3′ (SEQ ID NO:255) reversePCR primer 5′- CCACCTCAGGAAGCCGAAGATGCC-3′ (SEQ ID NO:256)

[1950] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA39621 sequence which had the followingnucleotide sequence:

[1951] hybridization probe

[1952] 5′-GAACGGTACAAGTGGCTGCGCTTCAGCGAGGACTGTCTGTACCTG-3′ (SEQ IDNO:257)

[1953] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO873 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB229).

[1954] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO873 [herein designated as UNQ440(DNA44179-1362)] (SEQ ID NO:253) and the derived protein sequence forPRO873.

[1955] The entire nucleotide sequence of UNQ440 (DNA44179-1362) is shownin FIG. 90 (SEQ ID NO:253). Clone UNQ440 (DNA44179-1362) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 139-141 and ending at the stop codon atnucleotide positions 1774-1776 (FIG. 90). The predicted polypeptideprecursor is 545 amino acids long (FIG. 91). The full-length PRO873protein shown in FIG. 91 has an estimated molecular weight of about58,934 daltons and a pI of about 9.45. Analysis of the full-lengthPRO873 sequence shown in FIG. 91 (SEQ ID NO:254) evidences the presenceof the following features: a signal peptide from about amino acid 1 toabout amino acid 29; a carboxylesterase type-B serine active site atabout amino acid 312 to about amino acid 327; a carboxylesterase type-Bsignature 2 motif at about amino acid 218 to about amino acid 228; andthree potential N-glycosylation sites at about amino acid 318 to aboutamino acid 321, about amino acid 380 to about amino acid 383, and aboutamino acid 465 to about amino acid 468. Clone UNQ440 (DNA44179-1362) hasbeen deposited with ATCC on May 6, 1998 and is assigned ATCC deposit no.209851.

[1956] Analysis of the amino acid sequence of the full-length PRO873polypeptide suggests that it possesses significant sequence similarityto a human liver carboxylesterase, thereby indicating that PRO873 may bea novel carboxylesterase. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO873 amino acid sequence and the following Dayhoffsequences: ES10_RAT, GEN12405, AB010633_(—)1, EST4_RAT, A48809,SASB_ANAPL, RNU41662_(—)1, RNU22952_(—)1, BAL_RAT, GEN13522.

Example 42 Isolation of cDNA Clones Encoding Human PRO940

[1957] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA47442. Based on the DNA47442consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO940.

[1958] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CAAAGCCTGCGCCTGGTCTGTG-3′ (SEQ ID NO:260) reversePCR primer 5′-TTCTGGAGCCCAGAGGGTGCTGAG-3′ (SEQ ID NO:262)

[1959] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA47442 sequence which had the followingnucleotide sequence

[1960] hybridization probe

[1961] 5′-GGAGCTGCCACCCATTCAAATGGAGCACGAAGGAGAGTTCACCTG-3′ (SEQ IDNO:263)

[1962] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was ii screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO940 gene usingthe probe oligonucleotide and one of the PCR primers. RNA e forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB229).

[1963] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO940 [herein designated as UNQ477(DNA54002-1367)] (SEQ ID NO:258) and the derived protein sequence forPRO940.

[1964] The entire nucleotide sequence of UNQ477 (DNA54002-1367) is shownin FIG. 92 (SEQ ID NO:258). Clone UNQ477 (DNA54002-1367) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 46-48 and ending at the stop codon at nucleotidepositions 1678-1680 (FIG. 92). The predicted polypeptide precursor is544 amino acids long (FIG. 93). The full-length PRO940 protein shown inFIG. 93 has an estimated molecular weight of about 60,268 daltons and apI of about 9.53. Analysis of the full-length PRO940 sequence shown inFIG. 93 (SEQ ID NO:259) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 15, potentialN-glycosylation sites from about amino acid 100 to about amino acid 103,from about amino acid 297 to about amino acid 300 and from about aminoacid 306 to about amino acid 309 and an immunoglobulin and majorhistocompatibility complex signature sequence block from about aminoacid 365 to about amino acid 371. Clone UNQ477 (DNA54002-1367) has beendeposited with ATCC on Apr. 7, 1998 and is assigned ATCC deposit no.209754.

[1965] Analysis of the amino acid sequence of the full-length PRO940polypeptide suggests that it possesses significant sequence similarityto CD33 and the OB binding protein-2. More specifically, an analysis ofthe Dayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO940 amino acid sequence and the followingDayhoff sequences, CD33_HUMAN, HSU71382_(—)1, HSU71383_(—)1,D86359_(—)1, PGBM_HUMAN, MAGS_MOUSE, D86983_(—)1, C22B_HUMAN, P_W01002and HVU24116_(—)1.

Example 43 Isolation of cDNA Clones Encoding Human PRO941

[1966] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA35941. An EST sequenceproprietary to Genentech was employed in the assembly and is hereindesignated DNA6415 (FIG. 96; SEQ ID NO:265). Based on the DNA35941consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO941.

[1967] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CTTGACTGTCTCTGAATCTGCACCC-3′ (SEQ ID NO:266)reverse PCR primer 5′-AAGTGGTGGAAGCCTCCAGTGTGG-3′ (SEQ ID NO:267)

[1968] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35941 sequence which had the followingnucleotide sequence

[1969] hybridization probe

[1970] 5′-CCACTACGGTATTAGAGCAAAAGTTAAAACCATCATGGTTCCTGGAGCAGC-3′ (SEQ IDNO:268)

[1971] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO941 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1972] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO941 [herein designated as UNQ478(DNA53906-1368)] (SEQ ID NO:263) and the derived protein sequence forPRO941.

[1973] The entire nucleotide sequence of UNQ478 (DNA53906-1368) is shownin FIG. 94 (SEQ ID NO:263). Clone UNQ478 (DNA53906-1368) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 37-39 and ending at the stop codon at nucleotidepositions 2353-2355 (FIG. 94). The predicted polypeptide precursor is772 amino acids long (FIG. 95). The full-length PRO941 protein shown inFIG. 95 has an estimated molecular weight of about 87,002 daltons and apI of about 4.64. Analysis of the full-length PRO941 sequence shown inFIG. 95 (SEQ ID NO:264) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 21, potentialN-glycosylation sites from about amino acid 57 to about amino acid 60,from about amino acid 74 to about amino acid 77, from about amino acid419 to about amino acid 422, from about amino acid 437 to about aminoacid 440, from about amino acid 508 to about amino acid 511, from aboutamino acid 515 to about amino acid 518, from about amino acid 516 toabout amino acid 519 and from about amino acid 534 to about amino acid537, and cadherin extracellular repeated domain signature sequences fromabout amino acid 136 to about amino acid 146 and from about amino acid244 to about amino acid 254. Clone UNQ478 (DNA53906-1368) has beendeposited with ATCC on Apr. 7, 1998 and is assigned ATCC deposit no.209747.

[1974] Analysis of the amino acid sequence of the full-length PRO941polypeptide suggests that it possesses significant sequence similarityto a cadherin protein, thereby indicating that PRO941 may be a novelcadherin protein family member. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO941 amino acid sequence and the followingDayhoff sequences, I50180, CADA_CHICK, I50178, GEN12782, CADC_HUMAN,P_W25637, A38992, P_R49731, D38992 and G02678.

Example 44 Isolation of cDNA Clones Encoding Human PRO944

[1975] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA47374. A variety ofproprietary Genentech EST sequences were employed in the assembly andare shown in FIGS. 99-107. Based on the DNA47374 consensus sequence,oligonucleotides were synthesized: 1) to identify by PCR a cDNA librarythat contained the sequence of interest, and 2) for use as probes toisolate a clone of the full-length coding sequence for PRO944.

[1976] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CGAGCGAGTCATGGCCAACGC-3′ (SEQ ID NO:280) reversePCR primer 5′-GTGTCACACGTAGTCTTTCCCGCTGG-3′ (SEQ ID NO:281)

[1977] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA47374 sequence which had the followingnucleotide sequence

[1978] hybridization probe

[1979] 5′-CTGCAGCTGTTGGGCTTCATTCTCGCCTTCCTGGGATGGATCG-3′ (SEQ ID NO:282)

[1980] In order to screen several libraries for a source of a fudllengthclone, DNA from the libraries was screened by PCR amplification with thePCR primer pair identified above. A positive library was then used toisolate clones encoding the PRO944 gene using the probe oligonucleotideand one of the PCR primers. RNA for construction of the cDNA librarieswas isolated from human fetal kidney tissue (LIB227).

[1981] DNA sequencing of the clones isolated as described above gavethne full-length DNA sequence for PRO944 herein designated as UNQ481 ()NA52185-1370)] (SEQ ID NO:269) and the derived protein sequence forPRO944.

[1982] The entire nucleotide sequence of UNQ481 (DNA52185-1370) is shownin FIG. 97 (SEQ ID NO:269). Clone UNQ481 (DNA52185-1370) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 219-221 and ending at the stop codon atnucleotide positions 852-854 (FIG. 97). The predicted polypeptideprecursor is 211 amino acids long (FIG. 98). The full-length PRO944protein shown in FIG. 98 has an estimated molecular weight of about22,744 daltons and a pI of about 8.51. Analysis of the full-lengthPRO944 sequence shown in FIG. 98 (SEQ ID NO:270) evidences the presenceof the following: a signal peptide from about amino acid 1 to aboutamino acid 21, transmembrane domains from about amino acid 82 to aboutamino acid 102, from about amino acid 118 to about amino acid 142 andfrom about amino acid 161 to about amino acid 187, a potentialN-glycosylation site from about amino acid 72 to about amino acid 757 asequence block having homology to PMP-22/EMP/MP20 family of proteinsfrom about amino acid 70 to about amino acid 111 and a sequence blockhaving homology to ABC-2 type transport system integral membrane proteinfrom about amino acid 119 to about amino acid 133. Clone UNQ481(DNA52185-1370) has been deposited with ATCC on May 14, 1998 and isassigned ATCC deposit no. 209861.

[1983] Analysis of the amino acid sequence of the full-length PRO944polypeptide suggests that it possesses significant sequence similarityto the CPE-R protein, thereby indicating that PRO944 may be a novelCPE-R homolog. More specifically, an analysis of the Dayhoff database(version 35.45 SwissProt 35) evidenced significant homology between thePRO944 amino acid sequence and the following Dayhoff sequences,AB000713_(—)1, AB000714_(—)1, AF035814_(—)1, AF000959_(—)1,HSU89916_(—)1, EMP2_HUMAN, JC5732, CELF53B3_(—)6, PM22_MOUSE andCGU49797_(—)1.

Example 45 Isolation of cDNA Clones Encoding Human PRO983

[1984] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA47473. Various proprietaryGenentech EST sequences were employed in the assembly, wherein those ESTsequences are shown in FIGS. 110-116. Based on the DNA47473 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence for PRO983.

[1985] A pair of PCR primners (forward and reverse) were synthesized:forward PCR primer 5′-GCACCACCGTAGGTACTTGTGTGAGGC-3′ (SEQ ID NO:292)reverse PCR primer 5′-AACCACCAGAGCCAAGAGCCGGG-3′ (SEQ ID NO:293)

[1986] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA47473 sequence which had the followingnucleotide sequence

[1987] hybridization probe

[1988] 5′-CAGCGGAATCATCGATGCAGGGGCCTCAATTAATGTATCTGTGATGTTAC-3′ (SEQ IDNO:294)

[1989] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO983 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrow(LIB256).

[1990] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO983 [herein designated as UNQ484(DNA53977-1371)] (SEQ ID NO:283) and the derived protein sequence forPRO983.

[1991] The entire nucleotide sequence of UNQ484 (DNA53977-1371) is shownin FIG. 108 (SEQ ID NO:283). Clone UNQ484 (DNA53977-1371) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 234-236 and ending at the stop codon atnucleotide positions 963-965 (FIG. 108). The predicted polypeptideprecursor is 243 amino acids long (FIG. 109). The full-length PRO983protein shown in FIG. 109 has an estimated molecular weight of about27,228 daltons and a pI of about 7.43. Analysis of the full-lengthPRO983 sequence shown in FIG. 109 (SEQ ID NO:284) evidences the presenceof the following features: a putative transmembrane domain from aboutamino acid 224 to about amino acid 239; a potential N-glycosylation sitefrom about amino acid 68 to about amino acid 71; and three potentialN-myristoylation sites from about amino acid 59 to about amino acid 64,from about amino acid 64 to about amino acid 69, and from about aminoacid 235 to about amino acid 240. Clone UNQ484 (DNA53977-1371) has beendeposited with ATCC on May 14, 1998 and is assigned ATCC deposit no.209862.

[1992] Analysis of the amino acid sequence of the full-length PRO983polypeptide suggests that it possesses significant sequence similarityto the vesicle-associated protein, VAP-33, thereby indicating thatPRO983 may be a novel vesicle associated membrane protein. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant homology between the PRO983 aminoacid sequence and the following Dayhoff sequences: VP33_APLCA,CELF33D11_(—)12, CELF42G2_(—)2, S50623, YDFC_SCHPO, CELF54H5_(—)2,CELZC196_(—)8, CEF57A10_(—)3, MSP3_GLORO, CEC15H1_(—)1.

Example 46 Isolation of cDNA Clones Encoding Human PRO1057

[1993] A consensus sequence was obtained relative to a variety of ESTsequences as described in Examnple 1 above, wherein the consensussequence obtained is herein designated DNA49808. Based on the DNA49808consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO1057.

[1994] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-GCATCTGCAGGAGAGAGCGAAGGG-3′ (SEQ ID NO:297) reverse PCR primer5′-CATCGTTCCCGTGAATCCAGAGGC-3′ (SEQ ID NO:298)

[1995] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA49808 sequence which had the followingnucleotide sequence

[1996] hybridization probe

[1997] 5′-GAAGGGAGGCCTTCCTTTCAGTGGACCCGGGTCAAGAATACCCAC-3′ (SEQ IDNO:299)

[1998] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1057 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[1999] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1057 [herein designated as UNQ522(DNA57253-1382)] (SEQ ID NO:295) and the derived protein sequence forPRO1057.

[2000] The entire nucleotide sequence of UNQ522 (DNA57253-1382) is shownin FIG. 117 (SEQ ID NO:295). Clone UNQ522 (DNA57253-1382) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 275-277 and ending at the stop codon atnucleotide positions 1514-1516 (FIG. 117). The predicted polypeptideprecursor is 413 amino acids long (FIG. 118). The full-length PRO1057protein shown in FIG. 118 has an estimated molecular weight of about47,070 daltons and a pI of about 9.92. Analysis of the full-lengthPRO1057 sequence shown in FIG. 118 (SEQ ID NO:296) evidences thepresence of the following: a signal peptide from about amino acid 1 toabout amino acid 16, potential N-glycosylation sites from about aminoacid 90 to about amino acid 93, from about amino acid 110 to about aminoacid 113 and from about amino acid 193 to about amino acid 196, aglycosaminoglycan attachment site from about amino acid 236 to aboutamino acid 239 and a serine protease histidine-containing active sitefrom about amino acid 165 to about amino acid 170. Clone UNQ522(DNA57253-1382) has been deposited with ATCC on May 14, 1998 and isassigned ATCC deposit no. 209867.

[2001] Analysis of the amino acid sequence of the full-length PRO1057polypeptide suggests that it possesses significant sequence similarityto various protease proteins, thereby indicating that PRO1057 may be anovel protease. More specifically, an analysis of the Dayhoff database(version 35.45 SwissProt 35) evidenced significant homology between thePRO1057 amino acid sequence and the following Dayhoff sequences,TRYE_DROER, P_R14159, A69660, EBN1_EBV, S65494, GEN12688, A51084_(—)1,P_R99571, A57514 and AF003200_(—)1.

Example 47 Isolation of cDNA Clones Encoding Human PRO1071

[2002] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA53035. Based on the DNA53035consensus sequence, it was determined that that consensus sequenceshared significant sequence identity with Incyte EST clone no. 2872569,a clone that upon review appeared to encode a full length protein. Assuch, Incyte EST clone no. 2872569 was purchased and its insert wasobtained and sequenced so as to confirm the proper sequence. Thissequence is herein designated UNQ528 or DNA58847-1383.

[2003] DNA sequencing of the clone isolated as described above gave thefull-length DNA sequence for PRO1071 [herein designated as UNQ528(DNA58847-1383)] (SEQ ID NO:300) and the derived protein sequence forPRO1071.

[2004] The entire nucleotide sequence of UNQ528 (DNA58847-1383) is shownin FIG. 119 (SEQ ID NO:300). Clone UNQ528 (DNA58848-1383) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 133-135 and ending at the stop codon atnucleotide positions 1708-1710 (FIG. 119). The predicted polypeptideprecursor is 525 amino acids long (FIG. 120). The full-length PRO1071protein shown in FIG. 120 has an estimated molecular weight of about58,416 daltons and a pI of about 6.62. Analysis of the full-lengthPRO1071 sequence shown in FIG. 120 (SEQ ID NO:301) evidences thepresence of the following: a signal peptide from about amino acid 1 toabout amino acid 25, a potential N- glycosylation site from about aminoacid 251 to about amino acid 254, a thrombospondin-1 homology block fromabout amino acid 385 to about amino acid 399 and von Willibrands factortype C homology blocks from about amino acid 385 to about amino acid399, from about amino acid 445 to about amino acid 459 and from aboutamino acid 42 to about amino acid 56. Clone UNQ528 (DNA58847-1383) hasbeen deposited with ATCC on May 20, 1998 and is assigned ATCC depositno. 209879.

[2005] Analysis of the amino acid sequence of the full-length PRO1071polypeptide suggests that it possesses significant sequence similarityto the thrombospondin protein, thereby indicating that PRO1071 may be anovel thrombospondin homolog. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO1071 amino acid sequence and the followingDayhoff sequences, AB002364_(—)1, D67076_(—)1, BTPCINPGN_(—)1,CET13H10_(—)1, CEF25H8_(—)5, CEF53B6_(—)2, CEC26C6_(—)6, HSSEMG_(—)1,CET21B6_(—)4 and BTY08561_(—)1.

Example 48 Isolation of cDNA Clones Encoding Human PRO1072

[2006] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA53125. Based on the DNA53125consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO1072.

[2007] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-CCAGGAAATGCTCCAGGAAGAGCC-3′ (SEQ ID NO:305) reverse PCR primer5′-GCCCATGACACCAAATTGAAGAGTGG-3′ (SEQ ID NO:306)

[2008] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA53125 sequence which had the followingnucleotide sequence

[2009] hybridization probe

[2010] 5′-AACGCAGGGATCTTCCAGTGCCCTTACATGAAGACTGAAGATGGG-3′ (SEQ IDNO:307)

[2011] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1072 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal lungtissue (LIB26).

[2012] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1072 [herein designated as UNQ529(DNA58747-1384)] (SEQ ID NO:302) and the derived protein sequence forPRO1072.

[2013] The entire nucleotide sequence of UNQ529 (DNA58747-1384) is shownin FIG. 121 (SEQ ID NO:302). Clone UNQ529 (DNA58747-1384) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 65-67 and ending at the stop codon at nucleotidepositions 1073-1075 (FIG. 121). The predicted polypeptide precursor is336 amino acids long (FIG. 122). The full-length PRO1072 protein shownin FIG. 122 has an estimated molecular weight of about 36,865 daltonsand a pI of about 9.15. Analysis of the full-length PRO1072 sequenceshown in FIG. 122 (SEQ ID NO:303) evidences the presence of thefollowing: a signal peptide from about amino acid 1 to about amino acid21, short-chain alcohol dehydrogenase protein homology blocks from aboutamino acid 134 to about amino acid 144, from about amino acid 44 toabout amino acid 56 and from about amino acid 239 to about amino acid248 and potential N-glycosylation sites from about amino acid 212 toabout amino acid 215 and from about amino acid 239 to about amino acid242. Clone UNQ529 (DNA58747-1384) has been deposited with ATCC on May14, 1998 and is assigned ATCC deposit no. 209868.

[2014] Analysis of the amino acid sequence of the full-length PRO1072polypeptide suggests that it possesses significant sequence similarityto the reductase family of proteins, thereby indicating that PRO1072 maybe a novel reductase. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO1072 amino acid sequence and the following Dayhoffsequences, P_W03198, P_W15759, P_R60800, MTV037_(—)3, CEC15H11_(—)6,ATAC00234314, MTV022_(—)13, SCU43704_(—)1, OXIR_STRAT AND CELC01G8_(—)3.

Example 49 Isolation of cDNA Clones Encoding Human PRO1075

[2015] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA34363. Based on the DNA34363sequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO1075.

[2016] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-TGAGAGGCCTCTCTGGAAGTTG-3′ (SEQ ID NO:312) forward PCR primer5′-GTCAGCGATCAGTGAAAGC-3′ (SEQ ID NO:313) forward PCR primer5′-CCAGAATGAAGTAGCTCGGC-3′ (SEQ ID NO:314) forward PCR primer5′-CCGACTCAAAATGCATTGTC-3′ (SEQ ID NO:315) forward PCR primer5′-CATTTGGCAGGAATTGTCC-3′ (SEQ ID NO:316) forward PCR primer5′-GGTGCTATAGGCCAAAGGG-3′ (SEQ ID NO:317) reverse PCR primer5′-CTGTATCTCTGGGCTATGTCAGAG-3′ (SEQ ID NO:318) reverse PCR primer5′-CTACATATAATGGCACATGTCAGCC-3′ (SEQ ID NO:319)

[2017] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA34363 sequence which had the followingnucleotide sequence

[2018] hybridization probe

[2019] 5′-CGTCTTCCTATCCTTACCCGACCTCAGATGCTCCCTTCTGCTCCTG-3′ (SEQ IDNO:320)

[2020] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1075 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human skin tumortissue (LIB324).

[2021] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1075 [herein designated as UNQ532(DNA57689-1385)] (SEQ ID NO:308) and the derived protein sequence forPRO1075.

[2022] The entire nucleotide sequence of UNQ532 (DNA57689-1385) is shownin FIG. 124 (SEQ ID NO:308). Clone UNQ532 (DNA57689-1385) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 137-139 and ending at the stop codon atnucleotide positions 1355-1357 (FIG. 124). The predicted polypeptideprecursor is 406 amino acids long (FIG. 125). The full-length PRO1075protein shown in FIG. 125 has an estimated molecular weight of about46,927 daltons and a pI of about 5.21. Analysis of the full-lengthPRO1075 sequence shown in FIG. 125 (SEQ ID NO:309) evidences thepresence of the following: a signal peptide from about amino acid 1 toabout amino acid 29, an endoplasmic reticulum targeting sequence fromabout amino acid 403 to about amino acid 406, a tyrosine kinasephosphorylation site from about amino acid 203 to about amino acid 211and a sequence block having homology to the thioredoxin family ofproteins from about amino acid 50 to about amino acid 66. Clone UNQ532(DNA57689-1385) has been deposited with ATCC on May 14, 1998 and isassigned ATCC deposit no. 209869.

[2023] Analysis of the amino acid sequence of the full-length PRO1075polypeptide suggests that it possesses significant sequence similarityto protein disulfide isomerase, thereby indicating that PRO1075 may be anovel protein disulfide isomerase. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO1075 amino acid sequence and the followingDayhoff sequences, CELC30H7_(—)2, CELC06A6_(—)3, CELF42G8_(—)3, S57942,ER72_CAEEL, CELC07A12_(—)3, CEH06O₀₁ _(—)4 and P_R51696.

Example 50 Isolation of cDNA Clones Encoding Human PRO181

[2024] A cDNA sequence isolated in the amylase screen described inExample 2 above was found, by BLAST and FastA sequence alignment, tohave sequence homology to a nucleotide sequence encoding the cornichonprotein. This cDNA sequence is herein designated DNA13242 (FIG. 130; SEQID NO:323). Based on the sequence homology, oligonucleotide probes weregenerated from the sequence of the DNA13242 molecule and used to screena human placenta (LIB89) library prepared as described in paragraph 1 ofExample 2 above. The cloning vector was pRK5B (pRK5B is a precursor ofpRK5D that does not contain the SfiI site; see, Holmes et al., Science,253:1278-1280 (1991)), and the cDNA size cut was less than 2800 bp.

[2025] The oligonucleotide probes employed included: forward PCR primer5′-GTGCAGCAGAGTGGCTTACA-3′ (SEQ ID NO:326) reverse PCR primer5′-ACTGGACCAATTCTTCTGTG-3′ (SEQ ID NO:327)

[2026] hybridization probe

[2027] 5′-GATATTCTAGCATATTGTCAGAAGGAAGGATGGTGCAAATTAGCT-3′ (SEQ IDNO:328)

[2028] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 14-16 and ending at the stop codon found atnucleotide positions 446-448 (FIG. 128; SEQ ID NO:321). The predictedpolypeptide precursor is 144 amino acids long, has a calculatedmolecular weight of approximately 16,699 daltons and an estimated pI ofapproximately 5.6. Analysis of the full-length PRO181 sequence shown inFIG. 129 (SEQ ID NO:322) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 20, aputative type II transmembrane domain from about amino acid 11 to aboutamino acid 31 and other transmembrane domains from about amino acid 57to about amino acid 77 and from about amino acid 123 to about amino acid143. Clone UNQ155 (DNA23330-1390) has been deposited with ATCC on Apr.14, 1998 and is assigned ATCC deposit no. 209775.

[2029] Analysis of the amino acid sequence of the full-length PRO181polypeptide suggests that it possesses significant sequence similarityto the cornichon protein, thereby indicating that PRO181 may be a novelcornichon homolog. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO181 amino acid sequence and the following Dayhoffsequences, AF022811_(—)1, CET09E8_(—)3, S64058, YGF4_YEAST, YB60_YEAST,EBU89455_(—)1, SIU36383_(—)3 and PH1371.

Example 51 Isolation of cDNA Clones Encoding Human PRO195

[2030] A cDNA sequence was isolated in the amylase screen described inExample 2 above and is herein designated DNA13199 (FIG. 134; SEQ IDNO:332). The DNA13199 sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) to identify existing homologies. The homologysearch was performed using the computer program BLAST or BLAST2 (Altshulet al., Methods in Enzymology 266:460-480 (1996)). Those comparisonsresulting in a BLAST score of 70 (or in some cases 90) or greater thatdid not encode known proteins were clustered and assembled intoconsensus DNA sequences with the program “phrap” (Phil Green, Universityof Washington, Seattle, Wash.;http://bozeman.mbt.washington.edu/phrap.docs/phrap.html). The consensussequence obtained therefrom is herein designated as DNA22778.

[2031] Based on the DNA22778 sequence, oligonucleotide probes weregenerated and used to screen a human placenta library (L1189) preparedas described in paragraph 1 of Example 2 above. The cloning vector waspRK5B (pRK5B is a precursor of pRK5D that does not contain the SfiIsite; see, Holmes et al., Science, 253:1278-1280 (1991)), and the cDNAsize cut was less than 2800 bp.

[2032] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-ACAAGCTGAGCTGCTGTGACAG-3′ (SEQ ID NO:333) reverse PCR primer5′-TGATTCTGGCAACCAAGATGGC-3′ (SEQ ID NO:334)

[2033] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA22778 sequence which had the followingnucleotide sequence

[2034] hybridization probe

[2035] 5′-ATGGCCTrGGCCGGAGGTTCGGGGACCGCTTCGGCTGAAG-3′ (SEQ ID NO:335)

[2036] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO195 gene usingthe probe oligonucleotide and one of the PCR primers.

[2037] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 70-72 and ending at the stop codon found atnucleotide positions 1039-1041 (FIG. 132; SEQ ID NO:330). The predictedpolypeptide precursor is 323 amino acids long, has a calculatedmolecular weight of approximately 36,223 daltons and an estimated pI ofapproximately 5.06. Analysis of the full-length PRO195 sequence shown inFIG. 132 (SEQ ID NO:330) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 31, atransmembrane domain from about amino acid 241 to about amino acid 260and a potential N-glycosylation site from about amino acid 90 to aboutamino acid 93. Clone UNQ169 (DNA26847-1395) has been deposited with ATCCon Apr. 14, 1998 and is assigned ATCC deposit no. 209772.

[2038] Analysis of the amino acid sequence of the full-length PRO195polypeptide suggests that it possesses no significant sequencesimilarity to any known protein. However, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced some degree of homologybetween the PRO195 amino acid sequence and the following Dayhoffsequences, P_P91380, AF035118_(—)1, HUMTROPCS_(—)1, NUOD_SALTY andE70002.

Example 52 Isolation of cDNA Clones Encoding Human PRO865

[2039] A cDNA sequence isolated in the amylase screen described inExample 2 above was herein designated DNA37642 (FIG. 137, SEQ IDNO:338). The DNA37642 sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ™,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify homologiestherebetween. The homology search was performed using the computerprogram BLAST or BLAST2 (Altshul et al., Methods in Enzymology266:460-480 (1996)). Those comparisons resulting in a BLAST score of 70(or in some cases 90) or greater that did not encode known proteins wereclustered and assembled into consensus DNA sequences with the program“phrap” (Phil Green, University of Washington, Seattle, Wash.;http://bozeman.mbt.washington.edulphrap.docs/phrap.html). The consensussequence obtained is herein designated DNA48615.

[2040] Based on the DNA48615 consensus sequence, probes were generatedand used to screen a human fetal kidney (LIB227) library prepared asdescribed in paragraph 1 of Example 2 above. The cloning vector waspRK5B (pRK5B is a precursor of pRK5D that does not contain the SfiIsite; see, Holmes et al., Science, 253:1278-1280 (1991)), and the cDNAsize cut was less than 2800 bp.

[2041] PCR primers (forward and reverse) were synthesized: forward PCRprimer 1 5′-AAGCTGCCGGAGCTGCAATG-3′ (SEQ ID NO:339) forward PCR primer 25′-TTGCTTCTTAATCCTGAGCGC-3′ (SEQ ID NO:340) forward PCR primer 35′-AAAGGAGGACTTTCGACTGC-3′ (SEQ ID NO:341) reverse PCR primer 15′-AGAGATTCATCCACTGCTCCAAGTCG-3′ (SEQ ID NO:342) reverse PCR primer 25′-TGTCCAGAAACAGGCACATATCAGC-3′ (SEQ ID NO:343)

[2042] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA48615 sequence which had the followingnucleotide sequence

[2043] hybridization probe

[2044] 5′-AGACAGCGGCACAGAGGTGCTTCTGCCAGGTTAGTGGTTACTTGGATGAT-3′ (SEQ IDNO:344)

[2045] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pairs identified above. A positivelibrary was then used to isolate clones encoding the PRO865 gene usingthe probe oligonucleotide and one of the PCR primers.

[2046] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 173-175 and ending at the stop codon found atnucleotide positions 1577-1579 (FIG. 135; SEQ ID NO:336). The predictedpolypeptide precursor is 468 amino acids long, has a calculatedmolecular weight of approximately 54,393 daltons and an estimated pI ofapproximately 5.63. Analysis of the full-length PRO865 sequence shown inFIG. 136 (SEQ ID NO:337) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 23, potentialN-glycosylation sites from about amino acid 280 to about amino acid 283and from about amino acid 384 to about amino acid 387, a potentialamidation site from about amino acid 94 to about amino acid 97,glycosaminoglycan attachment sites from about amino acid 20 to aboutamino acid 23 and from about amino acid 223 to about amino acid 226, anaminotransferase class-V pyridoxyl-phosphate amino acid sequence blockfrom about amino acid 216 to about amino acid 222 and an amino acidsequence block similar to that found in the interleukin-7 protein fromabout amino acid 338 to about amino acid 343. Clone UNQ434(DNA53974-1401) has been deposited with ATCC on Apr. 14, 1998 and isassigned ATCC deposit no. 209774.

[2047] Analysis of the amino acid sequence of the full-length PRO865polypeptide suggests that it possesses no significant sequencesimilarity to any known protein. However, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced some degree of homologybetween the PRO865 amino acid sequence and the following Dayhoffsequences, YMN0_YEAST, ATFCA4_(—)43, S44168, P_W14549 and RABTCRG4_(—)1.

Example 53 Isolation of cDNA Clones Encoding Human PRO827

[2048] A cDNA sequence isolated in the amylase screen described inExample 2 above was found, by BLAST and FastA sequence alignment, tohave sequence homology to nucleotide sequences encoding various integrinproteins. This cDNA sequence is herein designated DNA47751 (see FIG.140; SEQ ID NO:347). Based on the sequence homology, probes weregenerated from the sequence of the DNA47751 molecule and used to screena human fetal pigment epithelium library (LIB113) prepared as describedin paragraph 1 of Example 2 above. The cloning vector was pRK5B (pRK5Bis a precursor of pRK5D that does not contain the SfiI site; see, Holmeset al., Science 253:1278-1280 (1991)), and the cDNA size cut was lessthan 2800 bp.

[2049] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-AGGGACAGAGGCCAGAGGACTTC-3′ (SEQ ID NO:348) reverse PCR primer5′-CAGGTGCATATTCACAGCAGGATG-3′ (SEQ ID NO:349)

[2050] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA47751 sequence which had the followingnucleotide sequence

[2051] hybridization probe

[2052] 5′-GGAACTCCCCTTCGTCACTCACCTGTTCTTGCCCCTGGTGTTCCT-3′ (SEQ IDNO:350)

[2053] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO827 gene usingthe probe oligonucleotide and one of the PCR primers.

[2054] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 134-136 and ending at the stop codon found atnucleotide positions 506-508 (FIG. 138; SEQ ID NO:345). The predictedpolypeptide precursor is 124 amino acids long, has a calculatedmolecular weight of approximately 13,352 daltons and an estimated pI ofapproximately 5.99. Analysis of the full-length PRO827 sequence shown inFIG. 139 (SEQ ID NO:346) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 22, a cellattachment sequence from about amino acid 70 to about amino acid 72, apotential N-glycosylation site from about amino acid 98 to about aminoacid 101 and an integrin alpha chain protein homology sequence fromabout amino acid 67 to about amino acid 81. Clone UNQ468 (DNA57039-1402)has been deposited with ATCC on Apr. 14, 1998 and is assigned ATCCdeposit no. 209777.

[2055] Analysis of the amino acid sequence of the full-length PRO827polypeptide suggests that it possesses significant sequence similarityto the VLA-2 integrin protein and various other integrin proteins,thereby indicating that PRO827 may be a novel integrin or splice variantthereof. More specifically, an analysis of the Dayhoff database (version35.45 SwissProt 35) evidenced significant homology between the PRO240amino acid sequence and the following Dayhoff sequences, S44142,ITA2_HUMAN, ITA1_RAT, ITA1_HUMAN, ITA4_HUMAN, ITA9_HUMAN, AF032108_(—)1,ITAM_MOUSE, ITA8_CHICK and ITA6_CHICK.

Example 54 Isolation of cDNA Clones Encoding Human PRO1114

[2056] A cDNA sequence isolated in the amylase screen described inExample 2 was found, by the WU-BLAST2 sequence alignment computerprogram, to have certain sequence identity to other known interferonreceptors. This cDNA sequence is herein designated DNA48466 (FIG. 143;SEQ ID NO:352). Based on the sequence identity, probes were generatedfrom the sequence of the DNA48466 molecule and used to screen a humanbreast carconoma library (LIB135) prepared as described in paragraph 1of Example 2 above. The cloning vector was pRK5B (pRK5B is a precursorof pRK5D that does not contain the SfiI site; see, Holmes et al.,Science, 253:1278-1280 (1991)), and the cDNA size cut was less than 2800bp.

[2057] The oligonucleotide probes employed were as follows: forward PCRprimer 5′-AGGCTTCGCTGCGACTAGACCTC-3′ (SEQ ID NO:354) reverse PCR primer5′-CCAGGTCGGGTAAGGATGGTTGAG-3′ (SEQ ID NO:355)

[2058] hybridization probe

[2059] 5′-TTTCTACGCATTGATTCCATGTTTGCTCACAGATGAAGTGGCCATTCTGC-3′ (SEQ IDNO:356)

[2060] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 250-252, and a stop signal at nucleotide positions1183-1185 (FIG. 141, SEQ ID NO:351). The predicted polypeptide precursoris 311 amino acids long, has a calculated molecular weight ofapproximately 35,076 daltons and an estimated pI of approximately 5.04.Analysis of the full-length PRO1114 interferon receptor sequence shownin FIG. 142 (SEQ ID NO:352) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 29, atransmembrane domain from about amino acid 230 to about amino acid 255,potential N-glycosylation sites from about amino acid 40 to about aminoacid 43 and from about amino acid 134 to about amino acid 137, an aminoacid sequence block having homology to tissue factor proteins from aboutamino acid 92 to about amino acid 119 and an amino acid sequence blockhaving homology to integrin alpha chain proteins from about amino acid232 to about amino acid 262. Clone UNQ557 (DNA57033-1403) has beendeposited with ATCC on May 27, 1998 and is assigned ATCC deposit no.209905.

[2061] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 142 (SEQ ID NO:352), evidenced significanthomology between the PRO1114 interferon receptor amino acid sequence andthe following Dayhoff sequences: G01418, INR1_MOUSE, P_R71035,INGS_HUMAN, A26595_(—)1, A26593_(—)1, I56215 and TF_HUMAN.

Example 55 Isolation of cDNA Clones Encoding Human PRO237

[2062] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA30905. Based on the DNA30905consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO237.

[2063] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-TCTGCTGAGGTGCAGCTCATTCAC-3′ (SEQ ID NO:359) reverse PCR primer5′-GAGGCTCTGGAAGATCTGAGATGG-3′ (SEQ ID NO:360)

[2064] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA30905 sequence which had the followingnucleotide sequence

[2065] hybridization probe

[2066] 5′-GCCTCTTTGTCAACGTTGCCAGTACCTCTAACCCATTCCrCAGTCGCCTC-3′ (SEQ IDNO:361)

[2067] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO237 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal braintissue (L[B153).

[2068] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO237 [herein designated as UNQ211(DNA34353-1428)] (SEQ ID NO:357) and the derived protein sequence forPRO237.

[2069] The entire nucleotide sequence of UNQ211 (DNA34353-1428) is shownin FIG. 144 (SEQ ID NO:357). Clone UNQ211 (DNA34353-1428) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 586-588 and ending at the stop codon atnucleotide positions 1570-1572 (FIG. 144). The predicted polypeptideprecursor is 328 amino acids long (FIG. 145). The full-length PRO237protein shown in FIG. 145 has an estimated molecular weight of about36,238 daltons and a pI of about 9.90. Analysis of the full-lengthPRO237 sequence shown in FIG. 145 (SEQ ID NO:358) evidences the presenceof the following: a signal peptide from about amino acid 1 to aboutamino acid 23, a transmembrane domain from about amino acid 177 to aboutamino acid 199, potential N-glycosylation sites from about amino acid118 to about amino acid 121, from about amino acid 170 to about aminoacid 173 and from about amino acid 260 to about amino acid 263 andeukaryotic-type carbonic anhydrase sequence homology blocks from aboutamino acid 222 to about amino acid 270, from about amino acid 128 toabout amino acid 164 and from about amino acid 45 to about amino acid92. Clone UNQ211 (DNA34353-1428) has been deposited with ATCC on May 12,1998 and is assigned ATCC deposit no. 209855.

[2070] Analysis of the amino acid sequence of the full-length PRO237polypeptide suggests that it possesses significant sequence similarityto the carbonic anhydrase protein. More specifically, an analysis of theDayhoff database (version 35.45 SwissProt 35) evidenced significanthomology between the PRO237 amino acid sequence and the followingDayhoff sequences, AF050106_(—)1, OACALP_(—)1, CELD1022_(—)8,CAH2_HUMAN, 1CAC, CAH5_HUMAN, CAHP_HUMAN, CAH3_HUMAN, CAH1_HUMAN and2CAB.

Example 56 Isolation of cDNA Clones Encoding Human PRO541

[2071] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA42259. Based on the DNA42259consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO541.

[2072] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-GGACAGAATTTGGGAGCACACTGG-3′ (SEQ ID NO:364) forward PCR primer5′-CCAAGAGTATACTGTCCTCG-3′ (SEQ ID NO:365) reverse PCR primer5′-AGCACAGATTTTCTCTACAGCCCCC-3′ (SEQ ID NO:366) reverse PCR primer5′-AACCACTCCAGCATGTACTGCTGC-3′ (SEQ ID NO:367)

[2073] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA42259 sequence which had the followingnucleotide sequence

[2074] hybridization probe

[2075] 5′-CCATTCAGGTGTTCTGGCCCTGTATGTACACATTATACACAGGTCGTGTG-3′ (SEQ IDNO:368)

[2076] In order to screen several libraries for a source of a fudllengthclone, DNA from the libraries was screened by PCR amplification with oneof the PCR primer pairs identified above. A positive library was thenused to isolate clones encoding the PRO541 gene using the probeoligonucleotide and one of the PCR primers. RNA for construction of thecDNA libraries was isolated from human fetal kidney tissue (LIB227).

[2077] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO541 [herein designated as UNQ342(DNA45417-1432)] (SEQ ID NO:362) and the derived protein sequence forPRO541.

[2078] The entire nucleotide sequence of UNQ342 (DNA45417-1432) is shownin FIG. 146 (SEQ ID NO:362). Clone UNQ342 (DNA45417-1432) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 469-471 and ending at the stop codon atnucleotide positions 1969-1971 (FIG. 146). The predicted polypeptideprecursor is 500 amino acids long (FIG. 147). The full-length PRO541protein shown in FIG. 147 has an estimated molecular weight of about56,888 daltons and a pI of about 8.53. Analysis of the full-lengthPRO541 sequence shown in FIG. 147 (SEQ ID NO:363) evidences the presenceof the following: a signal peptide from about amino acid 1 to aboutamino acid 20, amino acid sequence blocks having homology toextracellular proteins SCP/Tpx-1/Ag5/PR-1/Sc7 from about amino acid 165to about amino acid 186, from about amino acid 196 to about amino acid218, from about amino acid 134 to about amino acid 146, from about aminoacid 96 to about amino acid 108 and from about amino acid 58 to aboutamino acid 77 and a potential N-glycosylation site from about amino acid28 to about amino acid 31. Clone UNQ342 (DNA45417-1432) has beendeposited with ATCC on May 27, 1998 and is assigned ATCC deposit no.209910.

[2079] Analysis of the amino acid sequence of the full-length PRO541polypeptide suggests that it possesses significant sequence similarityto a trypsin inhibitor protein, thereby indicating that PRO541 may be anovel trypsin inlhbitor. More specifically, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced significant homologybetween the PRO541 amino acid sequence and the following Dayhoffsequences, D45027_(—)1, AB009609_(—)1, JC5308, CRS3_HORSE, TPX1_HUMAN,HELO_HELHO, GEN14351, A28112_(—)1, CET05A10_(—)4 and P_W11485.

Example 57 Isolation of cDNA Clones Encoding Human PRO273

[2080] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA36465. Based on the DNA36465consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO273.

[2081] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CAGCGCCCTCCCCATGTCCCTG-3′ (SEQ ID NO:371) reversePCR primer 5′-TCCCAACTGGTTTGGAGTTTTCCC-3′ (SEQ ID NO:372)

[2082] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA36465 sequence which had the followingnucleotide sequence

[2083] hybridization probe

[2084] 5′-CTCCGGTCAGCATGAGGCTCCTGGCGGCCGCTGCTCCTGCTGCTG-3′ (SEQ IDNO:373)

[2085] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO273 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue.

[2086] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO273 [herein designated as UNQ240(DNA39523-1192)] (SEQ ID NO:369) and the derived protein sequence forPRO273.

[2087] The entire nucleotide sequence of UNQ240 (DNA39523-1192) is shownin FIG. 148 (SEQ ID NO:369). Clone UNQ240 (DNA39523-1192) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 167-169 and ending at the stop codon atnucleotide positions 500-502 (FIG. 148). The predicted polypeptideprecursor is 111 amino acids long (FIG. 149). Clone UNQ240(DNA39523-1192) has been deposited with the ATCC. It is understood thatthe deposited clone contains the actual sequence and that the sequencesprovided herein are merely representative based on current sequencingtechniques. Moreover, given the sequences provided herein and knowledgeof the universal genetic code, the corresponding nucleotides for anygiven amino acid can be routinely identified and vice versa.

[2088] Analysis of the amino acid sequence of the full-length PRO273polypeptide suggests that portions of it possess sequence identity withhuman macrophage inflammatory protein-2, cytokine-induced neutrophilchemoattractant 2, and neutrophil chemotactic factor 2-beta, therebyindicating that PRO273 is a novel chemokine.

[2089] As discussed further below, the cDNA was subcloned into abaculovirus vector and expressed in insect cells as a C-terminallytagged IgG fusion protein. N-terminal sequencing of the resultantprotein identified the signal sequence cleavage site, yielding a maturepolypeptide of 77 amino acids. The mature sequence, showing 31-40%identity to other human CXC chemokines, includes the four canonicalcysteine residues but lacks the ELR motif. Northern analysisdemonstrates expression at least in the small intestine, colon, spleen,lymph node and kidney. By in situ hybridization, also described indetail below, mRNA is localized to the lamina propria of intestinalvilli and to renal tubules.

Example 58 Isolation of cDNA Clones Encoding Human PRO701

[2090] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA39848. Based on the DNA39848consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO701.

[2091] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-GGCAAGCTACGGAAACGTCATCGTG-3′ (SEQ ID NO:376)reverse PCR primer 5′-AACCCCCGAGCCAAAAGATGGTCAC-3′ (SEQ ID NO:377)

[2092] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA39848 sequence which had the followingnucleotide sequence:

[2093] hybridization probe

[2094] 5′-GTACCGGTGACCAGGCAGCAAAAGGCAACTATGGGCTCCTGGATCAG-3′ (SEQ IDNO:378).

[2095] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO701 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[2096] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO701 [herein designated as UNQ365(DNA44205-1285)] (SEQ ID NO:374) and the derived protein sequence forPRO701.

[2097] The entire nucleotide sequence of UNQ365 (DNA44205-1285) is shownin FIG. 150 (SEQ ID NO:374). Clone UNQ365 (DNA44205-1285) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 50-52 and ending at the stop codon at nucleotidepositions 2498-3000 (FIG. 150). The predicted polypeptide precursor is816 amino acids long (FIG. 151). The full-length PRO701 protein shown inFIG. 151 has an estimated molecular weight of about 91,794 daltons, a pIof about 5.88 and NX(S/T) being 4. Clone UNQ365 (DNA44205-1285) has beendeposited with the ATCC on Mar. 31, 1998. It is understood that theclone was the correct and actual sequence, wherein the sequencesprovided herein are representative based on sequencing techniques.

[2098] Still regarding the amino acid sequence shown in FIG. 151, thereis a potential signal peptide cleavage site at about amino acid 25.There are potential N-glycosylation sites at about amino acid positions83, 511, 716 and 803. The carboxylesterases type-B signature 2 sequenceis at about residues 125 to 135. Regions homologous withcarboxylesterase type-B are also at about residues 54-74, 197-212 and221-261. A potential transmembrane region corresponds approximately toamino acids 671 through about 700. The corresponding nucleic acids canbe routinely determined from the sequences provided herein.

[2099] Analysis of the amino acid sequence of the full-length PRO701polypeptide suggests that it possess significant homology to theneuroligins from rattus norvegicus indicating that PRO701 may be a novelhuman neuroligin.

Example 59 Isolation of cDNA Clones Encoding Human PRO704

[2100] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43033. Based on the DNA43033consensus sequence, oligonucleotides were synthesized: 1) to identity byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO704.

[2101] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CCTTGGGTCGTGGCAGCAGTGG-3′; (SEQ ID NO:381) reversePCR primer 5′-CACTCTCCAGGCTGCATGCTCAGG-3′. (SEQ ID NO:382)

[2102] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA43033 consensus sequence which had the followingnucleotide sequence:

[2103] hybridization probe

[2104] 5′-GTCAAACGTTCGAGTACTTGAAACGGGAGCACTCGCTGTCGAAGC-3′ (SEQ IDNO:383).

[2105] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO704 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[2106] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO704 [herein designated as UNQ368(DNA50911-1288)] (SEQ ID NO:379) and the derived protein sequence forPRO704.

[2107] The entire nucleotide sequence of UNQ368 (DNA50911-1288) is shownin FIG. 152 (SEQ ID NO:379). Clone UNQ368 (DNA50911-1288) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 8-10 and ending at the stop codon at nucleotidepositions 1052-1054 (FIG. 152). The predicted polypeptide precursor is348 amino acids long (FIG. 153). The full-length PRO704 protein shown inFIG. 153 has an estimated molecular weight of about 39,711 and a pI ofabout 8.7. Clone UNQ368 (DNA50911-1288) has been deposited with the ATCCon Mar. 31, 1998. Regarding the sequence, it is understood that thedeposited clone contains the correct sequence, and the sequencesprovided herein are based on known sequencing techniques.

[2108] Analysis of the amino acid sequence of the full-length PRO704polypeptide suggests that portions of it possess significant homology tothe vesicular integral membrane protein 36, thereby indicating thatPRO704 may be a novel vesicular integral membrane protein.

[2109] Still analyzing the amino acid sequence of SEQ ID NO:380, theputative signal peptide is at about amino acids 1-39 of SEQ ID NO:380.The transmembrane domain is at amino acids 310-335 of SEQ ID NO:380. Apotential N-glycosylation site is at about amino acids 180-183 of SEQ IDNO:380. The corresponding nucleotides can be routinely determined giventhe sequences provided herein.

Example 60 Isolation of cDNA Clones Encoding Human PRO706

[2110] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA40669. Based on the DNA40669consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO706.

[2111] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CCAAGCAGCTTAGAGCTCCAGACC-3′ (SEQ ID NO:386)reverse PCR primer 5′-TTCCCTATGCTCTGTATTGGCATGG-3′ (SEQ ID NO:387)

[2112] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA40669 sequence which had the followingnucleotide sequence

[2113] hybridization probe

[2114] 5′-GCCACTTCTGCCACAATGTCAGCTTTCCCTGTACCAGAAATGGCTGTGTT-3′ (SEQ IDNO:388)

[2115] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO706 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal braintissue (LIB153).

[2116] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO706 therein designated as UNQ370(DNA48329-1290)] (SEQ ID NO:384) and the derived protein sequence forPRO706. It is understood that the deposited clone contains the actualsequence, and that the sequences provided herein are representativebased on current sequencing techniques.

[2117] The entire nucleotide sequence of UNQ370 (DNA48329-1290) is shownin FIG. 154 (SEQ ID NO:384). Clone UNQ370 (DNA48329-1290) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 279-281 and ending at the stop codon atnucleotide positions 1719-1721 (FIG. 154). The predicted polypeptideprecursor is 480 amino acids long (FIG. 155). The full-length PRO706protein shown in FIG. 155 has an estimated molecular weight of about55,239 daltons and a pI of about 9.30. Clone UNQ370 (DNA48329-1290) hasbeen deposited with the ATCC on Apr. 21, 1998.

[2118] Still regarding the amino acid sequence shown in FIG. 155, thereis a potential signal peptide cleavage site at about amino acid 19.There are potential N-glycosylation sites at about amino acid positions305 and 354. There is a potential tyrosine kinase phosphorylation siteat about amino acid position 333. A region homologous with histidineacid phosphatase is at about residues 87-102. The corresponding nucleicacid regions can be routinely determined given the provided sequences,i.e., the codons can be determined from the specifically named aminoacids given.

[2119] Analysis of the amino acid sequence of the full-length PRO706polypeptide suggests that portions of it possess significant homology tothe human prostatic acid phosphatase precursor thereby indicating thatPRO706 may be a novel human prostatic acid phosphatase.

Example 61 Isolation of cDNA Clones Encoding Human PRO707

[2120] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA42775. Based on DNA42775,oligonucleotides were synthesized: 1) to identify by PCR a cDNA librarythat contained the sequence of interest, and 2) for use as probes toisolate a clone of the full-length coding sequence for PRO707.

[2121] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-TCCGTCTCTGTGAACCGCCCCAC-3′; (SEQ ID NO:391)reverse PCR primer 5′-CTCGGGCGCATTGTCGTTCTGGTC-3′. (SEQ ID NO:392)

[2122] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA42775 sequence which had the followingnucleotide sequence:

[2123] hybridization probe

[2124] 5′-CCGACTGTGAAAGAGAACGCCCCAGATCCACTTATTCCCC-3′ (SEQ ID NO:393).

[2125] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO707 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[2126] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO707 [herein designated as UNQ371(DNA48306-1291)] (SEQ ID NO:389) and the derived protein sequence forPRO707.

[2127] The entire nucleotide sequence of UNQ371 (DNA48306-1291) is shownin FIG. 156 (SEQ ID NO:389). Clone UNQ371 (DNA48306-1291) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 371-373 and ending at the stop codon atnucleotide positions 3119-3121 of SEQ ID NO:389. The predictedpolypeptide precursor is 916 amino acids long (FIG. 157). Thefull-length PRO707 protein shown in FIG. 157 has an estimated molecularweight of about 100,204 daltons and a pI of about 4.92. Clone UNQ371(DNA48306-1291) has been deposited with ATCC on May 27, 1998. It isunderstood that the clone UNQ371 which is deposited is that whichencodes PRO707, and that the sequences herein are merely representationsbased on known sequencing techniques which may be subject to minorerrors.

[2128] Regarding analysis of the amino acid sequence, the signalsequence appears to be at about 1 through 30 of SEQ ID NO:390. Cadherinsextracellular repeated domain signature sequence is at about amino acids121-131, 230-240, 335-345, 440-450, and 550-560 of SEQ ID NO:390.Tyrosine kinase phosphorylation sites are at about amino acids 124-132and 580-586 of SEQ ID NO:390. A potential transmembrane domain is atabout amino acids 682-715. The nucleic acid positions can be derived byreferring to the corresponding codon for the named amino acid.

[2129] Analysis of the amino acid sequence of the full-length PRO707polypeptide suggests that portions of it possess significant homology tothe cadherin FIB3 protein, expressed in human fibroblasts, therebyindicating that PRO707 may be a novel cadherin.

Example 62 Isolation of cDNA Clones Encoding Human PRO322

[2130] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA48336. Based on the DNA48336consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO322.

[2131] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CAGCCTACAGAATAAAGATGGCCC-3′ (SEQ ID NO:396)reverse PCR primer 5′-GGTGCAATGATCTGCCAGGCTGAT-3′ (SEQ ID NO:397)

[2132] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA48336 consensus sequence which had the followingnucleotide sequence:

[2133] hybridization probe

[2134] 5′-AGAAATACCTGTGGTTCAGTCCATCCCAAACCCCTGCTACAACAGCAG-3′ (SEQ IDNO:398).

[2135] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO322 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[2136] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO322 [herein designated as UNQ283(DNA48336-1309)] (SEQ ID NO:394) and the derived protein sequence forPRO322. It is understood that UNQ283 (DNA48336-1309) in fact encodesPRO322, and that SEQ ID NO:394 is a representation of the sequence basedon sequencing techniques known in the art.

[2137] The entire nucleotide sequence of UNQ283 (DNA48336-1309) is shownin FIG. 158 (SEQ ID NO:394). Clone UNQ283 (DNA48336-1309) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 166-168 and ending at the stop codon atnucleotide positions 946-948 (FIG. 158). The predicted polypeptideprecursor is 260 amino acids long (FIG. 159). The full-length PRO322protein shown in FIG. 159 has an estimated molecular weight of about28,028 daltons and a pI of about 7.87. Clone UNQ283 (DNA48336-1309) hasbeen deposited with ATCC and is assigned ATCC deposit no. 209669.

[2138] Regarding the amino acid sequence of FIG. 159, a potentialN-glycosylation site is at amino acid 110 of SEQ ID NO:395. The serineproteases, trypsin family and histidine active site is identified atamino acids 69 through 74 of SEQ ID NO:395 and the consensus sequence isidentified at amino acids 207 through 217 of SEQ ID NO:395. The kringledomain proteins motif is identified at amino acids 205 through 217 ofSEQ ID NO:395. The putative signal peptide is encoded at about aminoacids 1-23.

[2139] Analysis of the amino acid sequence of the full-length PRO322polypeptide suggests that portions of it possess significant homology toneuropsin and other serine proteases, thereby indicating that PRO322 isa novel serine protease related to neuropsin.

Example 63 Isolation of cDNA Clones Encoding Human PRO526

[2140] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA39626. Based on the DNA39626consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO526.

[2141] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-TGGCTGCCCTGCAGTACCTCTACC-3′; (SEQ ID NO:401)reverse PCR primer 5′-CCCTGCAGGTCATTGGCAGCTAGG-3′. (SEQ ID NO:402)

[2142] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA39626 consensus sequence which had the followingnucleotide sequence:

[2143] hybridization probe

[2144] 5′-AGGCACTGCCTGATGACACCJTCCGCGACCTGGGCAACCTCACAC-3′ (SEQ IDNO:403).

[2145] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO526 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB228).

[2146] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO526 [herein designated as UNQ330(DNA44184-1319)] (SEQ ID NO:399) and the derived protein sequence forPRO526.

[2147] The entire nucleotide sequence of UNQ330 (DNA44184-1319) is shownin FIG. 160 (SEQ ID NO:399). Clone UNQ330 (DNA44184-1319) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 514-516 and ending at the stop codon atnucleotide positions 1933-1935 (FIG. 160). The predicted polypeptideprecursor is 473 amino acids long (FIG. 161). The full-length PRO526protein shown in FIG. 161 has an estimated molecular weight of about50,708 daltons and a pI of about 9.28. Clone UNQ330 (DNA44185-1319) hasbeen deposited with the ATCC on Mar. 26, 1998. It is understood that theclone contains the actual sequence, whereas the sequences presentedherein are representative based on current sequencing techniques.

[2148] Analysis of the amino acid sequence of the full-length PRO526polypeptide suggests that portions of it possess significant homology tothe leucine repeat rich proteins including ALS, SLIT, carboxypeptidaseand platelet glycoprotein V thereby indicating that PRO526 is a novelprotein which is involved in protein-protein interactions.

[2149] Still analyzing SEQ ID NO:400, the signal peptide sequence is atabout amino acids 1-26. A leucine zipper pattern is at about amino acids135-156. A glycosaminoglycan attachment is at about amino acids 436-439.N-glycosylation sites are at about amino acids 82-85, 179-182, 237-240and 423-426. A von Willebrand factor (VWF) type C domain(s) is found atabout amino acids 411-425. The skilled artisan can understand whichnucleotides correspond to these amino acids based on the sequencesprovided herein.

Example 64 Isolation of cDNA Clones Encoding Human PRO531

[2150] An ECD database was searched and an expressed sequence tag (EST)from LIFESEQ™, Incyte Pharmaceuticals, Palo Alto, Calif. was identifiedwhich showed homology to protocadherin 3. Based on this sequence, asearch was performed using the computer program BLAST or BLAST2 (Altshulet al., Methods in Enzymolozy 266:460-480 (1996)) as a comparison of theECD protein sequences to a 6 frame translation of the EST sequence.Those comparisons resulting in a BLAST score of 70 (or in some cases 90)or greater that did not encode known proteins were clustered andassembled into consensus DNA sequences with the program “phrap” (PhilGreen, University of Washington, Seattle, Wash.).

[2151] A consensus DNA sequence was assembled relative to other ESTsequences using phrap. Based on the consensus sequence obtained,oligonucleotides were synthesized: 1) to identify by PCR a cDNA librarythat contained the sequence of interest, and 2) for use as probes toisolate a clone of the full-length coding sequence for PRO531.

[2152] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CTGAGAACGCGCCTGAAACTGTG-3′; (SEQ ID NO:406)reverse PCR primer 5′-AGCGTTGTCATTGACATCGGCG-3′. (SEQ ID NO:407)

[2153] Additionally, a synthetic oligonucleotide hybridizationprobe wasconstructed from the consensus DNA sequence which had the followingnucleotide sequence:

[2154] hybridization probe

[2155] 5′-TTAGTTGCTCCATTCAGGAGGATCTACCCTTCCTCCTGAAATCCGCGGAA-3′ (SEQ IDNO:408).

[2156] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO531 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal braintissue (LIB 153). The cDNA libraries used to isolate the cDNA cloneswere constructed by standard methods using commercially availablereagents such as those from Invitrogen, San Diego, Calif. The cDNA wasprimed with oligo dT containing a NotI site, linked with blunt to Sailhemikinased adaptors, cleaved with NotI, sized appropriately by gelelectrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the SfiI site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

[2157] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO531 [herein designated as UNQ332(DNA48314-1320)] (SEQ ID NO:404) and the derived protein sequence forPRO531.

[2158] The entire representative nucleotide sequence of UNQ332(DNA48314-1320) is shown in FIG. 162 (SEQ ID NO:404). It is understoodthat the actual sequence is that within the clone deposited with theATCC as DNA48314-1320. Clone UNQ332 (DNA48314-1320) contains a singleopen reading frame with an apparent translational initiation site atnucleotide positions 171-173 and ending at the stop codon at nucleotidepositions 2565-2567 (FIG. 162). The predicted polypeptide precursor is789 amino acids long (FIG. 163). The full-length PRO531 protein shown inFIG. 163 has an estimated molecular weight of about 87,552 daltons and apI of about 4.84. Clone UNQ332 (DNA48314-1320) has been deposited withthe ATCC on Mar. 26, 1998.

[2159] Analysis of the amino acid sequence of the full-length PRO531polypeptide suggests that portions of it possess significant homology toprotocadherin 3. Moreover, PRO531 is found in the brain, like otherprotocadherins, thereby indicating that PRO531 is a novel member of thecadherin superfamily.

[2160] Still analyzing the amino acid sequence of SEQ ID NO:405, thecadherin extracellular repeated domain signature is found at about aminoacids 122-132, 231-241, 336-346, 439-449 and 549-559 of SEQ ID NO:405.An ATP/GTP-binding site motif A (P-loop) is found at about amino acids285-292 of SEQ ID NO:405. N-glycosylation sites are found at least atabout amino acids 567-570, 786-790, 418-421 and 336-339 of SEQ IDNO:405. The signal peptide is at about amino acids 1-26, and thetransmembrane domain is at about amino acids 685-712 of SEQ ID NO:405.

Example 65 Isolation of cDNA Clones Encoding Human PRO534

[2161] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43038. Based on the 43048consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO534.

[2162] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CACAGAGCCAGAAGTGGCGGAATC-3′; (SEQ ID NO:411)reverse PCR primer 5′-CCACATGTTCCTGCTCTTGTCCTGG-3′. (SEQ ID NO:412)

[2163] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA43038 sequence which had the followingnucleotide sequence:

[2164] hybridization probe

[2165] 5′-CGGTAGTGACTGTACTCTAGTCCTGTITACACCCCGTGGTGCCG-3′ (SEQ IDNO:413).

[2166] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO534 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal lungtissue (LIB26).

[2167] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO534 [herein designated as UNQ335(DNA48333-1321)] (SEQ ID NO:409) and the derived protein sequence forPRO534.

[2168] The entire nucleotide sequence of UNQ335 (DNA48333-1321) is shownin FIG. 164 (SEQ ID NO:409). Clone UNQ335 (DNA48333-1321) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 87-89 and ending at the stop codon at nucleotidepositions 1167-1169 (FIG. 164). The predicted polypeptide precursor is360 amino acids long (FIG. 165). The full-length PRO534 protein shown inFIG. 165 has an estimated molecular weight of about 39,885 daltons and apI of about 4.79. Clone UNQ335 (DNA48333-1321) has been deposited withATCC on Mar. 26, 1998. It is understood that the deposited clonecontains the actual sequence, and that the sequences provided herein arerepresentative based on current sequencing techniques.

[2169] Analysis of the amino acid sequence of the full-length PRO534polypeptide suggests that portions of it possess significant sequenceidentity with the protein disulfide isomerase, thereby indicating thatPRO534 may be a novel disulfide isomerase.

[2170] Still analyzing the amino acid sequence of PRO534, the signalpeptides is at about amino acids 1-25 of SEQ ID NO:410. Thetransmembrane domain is at about amino acids 321-340 of SEQ ID NO:410.The disulfide isomerase corresponding region is at amino acids 212-302of SEQ ID NO:410. The thioredoxin domain is at amino acids 211-227 ofSEQ ID NO:410. N-glycosylation sites are at: 165-168, 181-184, 187-190,194-197, 206-209, 278-281, and 293-296 of SEQ ID NO:410. Thecorresponding nucleotides can routinely be determined from the sequencesprovided herein. PRO534 has a transmembrane domain rather than an ERretention peptide like other protein disulfide isomerases. Additionally,PRO534 may have an intron at the 5 prime end.

Example 66 Isolation of cDNA Clones Encoding Human PRO697

[2171] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43052. Based on this consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence for PRO697.

[2172] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CCTGGCTCGCTGCTGCTGCTC-3′; (SEQ ID NO:416) reversePCR primer 5′-CCTCACAGGTGCACTGCAAGCTGTC-3′. (SEQ ID NO:417)

[2173] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA43052 consensus sequence which had the followingnucleotide sequence:

[2174] hybridization probe

[2175] 5′-CTCTTCCTCTTTGGCCAGCCCGACTTCTCCTACAAGCGCAGAATTGC-3′ (SEQ IDNO:418).

[2176] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO697 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[2177] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO697 [herein designated as UNQ361(DNA50920-1325)] (SEQ ID NO:414) and the derived protein sequence forPRO697.

[2178] The entire nucleotide sequence of UNQ361 (DNA50920-1325) is shownin FIG. 166 (SEQ ID NO:414). Clone UNQ361 (DNA50920-1325) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 4446 and ending at the stop codon at nucleotidepositions 929-931 (FIG. 166). The predicted polypeptide precursor is 295amino acids long (FIG. 167). The full-length PRO697 protein shown inFIG. 167 has an estimated molecular weight of about 33,518 daltons and apI of about 7.74. Clone UNQ361 (DNA50920-1325) was deposited with theATCC on Mar. 26, 1998. It is understood that the deposited clonecontains the actual sequence, and that the sequences provided herein arerepresentative based on current sequencing techniques.

[2179] Analysis of the amino acid sequence of the full-length PRO697polypeptide suggests that portions of it possess significant sequenceidentity with sFRPs, thereby indicating that PRO697 may be a novel sFRPfamily member.

[2180] Still analyzing the amino acid sequence of PRO697, the signalpeptides is at about amino acids 1-20 of SEQ ID NO:415. The cystein richdomain, having identity with the frizzled N-terminus, is at about aminoacids 153 of SEQ ID NO:415. The corresponding nucleotides can routinelybe determined from the sequences provided herein.

Example 67 Isolation of cDNA Clones Encoding Human PRO717

[2181] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA42829. Based on the DNA42829consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO717.

[2182] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-AGCTTCTCAGCCCTCCTGGAGCAG-3′; (SEQ ID NO:421)reverse PCR primer 5′-CGGGTCAATAAACCTGGACGCTTGG-3′. (SEQ ID NO:422)

[2183] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA42829 consensus sequence which had the followingnucleotide sequence:

[2184] hybridization probe

[2185] 5′-TATGTGGACCGGACCAAGCACTTCACTGAGGCCACCAAGATTG-3′ (SEQ IDNO:423).

[2186] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO717 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal livertissue (LIB229).

[2187] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO717 [herein designated as UNQ385(DNA50988-1326)] (SEQ ID NO:419) and the derived protein sequence forPRO717.

[2188] The entire nucleotide sequence of UNQ385 (DNA50988-1326) is shownin FIG. 168 (SEQ ID NO:419). Clone UNQ385 (DNA50988-1326) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 17-19 and ending at the stop codon at nucleotidepositions 1697-1699 (FIG. 168). The predicted polypeptide precursor is560 amino acids long (FIG. 169). The full-length PRO717 protein shown inFIG. 169 has an estimated molecular weight of about 58,427 daltons and apI of about 6.86. Clone UNQ385 (DNA50988-1326) has been deposited withthe ATCC on Apr. 28, 1998. Regarding the sequence, it is understood thatthe deposited clone contains the correct sequence, and the ;sequencesprovided herein are based on known sequencing techniques.

[2189] Analysis of the amino acid sequence of the full-length PRO717polypeptide suggests that PRO717 may be a novel 12 transmembranereceptor. The reverse complement strand of DNA50988 has a stretch thatmatches identically with human regulatory myosin light strand.

[2190] Still analyzing the amino acid sequence of SEQ ID NO:420,transmembrane domains are at about amino acids 30-50, 61-79, 98-112,126-146, 169-182, 201-215, 248-268, 280-300, 318-337, 341-357, 375-387,and 420-441 of SEQ ID NO:420. N-glycosylation sites are at about aminoacids 40-43 and 4346 of SEQ ID NO:420. A glycosaminoglycan attachmentsite is at about amino acids 468-471 of SEQ ID NO:420. The correspondingnucleotides can be routinely determined given the sequences providedherein.

Example 68 Isolation of cDNA Clones Encoding Human PRO731

[2191] A database was used to search expressed sequence tag (EST)databases. The EST database used herein was the proprietary EST DNAdatabase LIFESEQ™, of Incyte Pharmaceuticals, Palo Alto, Calif. Incyteclone 2581326 was herein identified and termed DNA42801. Based on theDNA42801 sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO731.

[2192] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-GTAAGCACATGCCTCCAGAGGTGC-3′; (SEQ ID NO:426)reverse PCR primer 5′-GTGACGTGGATGCTTGGGATGTTG-3′. (SEQ ID NO:427)

[2193] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA42801 sequence which had the followingnucleotide sequence:

[2194] hybridization probe

[2195] 5′-TGGACACCTTCAGTATTGATGCCAAGACAGGCCAGGTCATTCTGCGTCGA-3′ (SEQ IDNO:428).

[2196] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO731 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrowtissue (LIB255). The cDNA libraries used to isolate the cDNA clones wereconstructed by standard methods using commercially available reagentssuch as those from Invitrogen, San Diego, Calif. The cDNA was primedwith oligo dT containing a NotI site, linked with blunt to SalIhemikinased adaptors, cleaved with NotI, sized appropriately by gelelectrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the Sfil site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

[2197] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO731 [herein designated as UNQ395(DNA48331-1329)] (SEQ ID NO:424) and the derived protein sequence forPRO731.

[2198] The entire nucleotide sequence of UNQ395 (DNA48331-1329) is shownin FIG. 170 (SEQ ID NO:424). Clone UNQ395 (DNA48331-1329) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 329-331 and ending at the stop codon atnucleotide positions 3881-3883 (FIG. 170). The predicted polypeptideprecursor is 1184 amino acids long (FIG. 171). The full-length PRO731protein shown in FIG. 171 has an estimated molecular weight of about129,022 daltons and a pI of about 5.2. Clone UNQ395 (DNA48331-1329) wasdeposited with the ATCC on Mar. 31, 1998. Regarding the sequence, it isunderstood that the deposited clone contains the correct sequence, andthe sequences provided herein are based on known sequencing techniques.

[2199] Analysis of the amino acid sequence of the full-length PRO731polypeptide suggests that portions of it possess significant identityand similarity to members of the protocadherin family, therebyindicating that PRO731 may be a novel protocadherin.

[2200] Still analyzing the amino acid sequence of SEQ ID NO:425, theputative signal peptide is at about amino acids 1-13 of SEQ ID NO:425.The transmembrane domain is at amino acids 719-739 of SEQ ID NO:425. TheN-glycosylation of SEQ ID NO:425 are as follows: 415-418, 582-586,659-662, 662-665, and 857-860. The cadherin extracellular repeateddomain signatures are at about amino acids (of SEQ ID NO:425): 123-133,232-242, 340-350, 448-458, and 553-563. The corresponding nucleotidescan be routinely determined given the sequences provided herein.

Example 69 Isolation of cDNA Clones Encoding Human PRO218

[2201] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA17411. Two proprietaryGenentech EST sequences were employed in the consensus assembly and areshown in FIGS. 174 and 175. Based on the DNA17411 consensus sequence,oligonucleotides were synthesized: 1) to identify by PCR a cDNA librarythat contained the sequence of interest, and 2) for use as probes toisolate a clone of the full-length coding sequence for PRO218.

[2202] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-AAGTGGAGCCGGAGCCTTCC-3′; (SEQ ID NO:433) reversePCR primer 5′-TCGTTGTTTATGCAGTAGTCGG-3′. (SEQ ID NO:434)

[2203] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA17411 sequence which had the followingnucleotide sequence:

[2204] hybridization probe

[2205] 5′-ATTGTTTAAAGACTATGAGATACGTCAGTATGTTGTACAGG-3′ (SEQ ID NO:435).

[2206] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO218 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB28).

[2207] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO218 [herein designated as UNQ192(DNA30867-1335)] (SEQ ID NO:429) and the derived protein sequence forPRO218.

[2208] The entire nucleotide sequence of UNQ192 (DNA30867-1335) is shownin FIG. 172 (SEQ ID NO:429). Clone UNQ192 (DNA30867-1335) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 150-152 and ending at the stop codon atnucleotide positions 1515-1517 (FIG. 172). The predicted polypeptideprecursor is 455 amino acids long (FIG. 173). The full-length PRO218protein shown in FIG. 173 has an estimated molecular weight of about52,917 daltons and a pI of about 9.5. Clone UNQ192 (DNA30867-1335) hasbeen deposited with the ATCC on Apr. 28, 1998. Regarding the sequence,it is understood that the deposited clone contains the correct sequence,and the sequences provided herein are based on known sequencingtechniques.

[2209] Analysis of the amino acid sequence of the full-length PRO218polypeptide suggests that PRO218 may be a novel trausmembrane protein.

[2210] Still analyzing the amino acid sequence of SEQ ID NO:430, theputative signal peptide is at about amino acids 1 through 23 of SEQ IDNO:430. Transmembrane domains are potentially at about amino acids37-55, 81-102, 150-168, 288-311, 338-356, 375-398, and 425-444 of SEQ IDNO:430. N-glycosylation sites are at about amino acids 67, 180, and 243of SEQ ID NO:430. Eukaryotic cobalamin-binding protein is at about aminoacids 151-160 of SEQ ID NO:430. The corresponding nucleotides can beroutinely determined given the sequences provided herein.

Example 70 Isolation of cDNA Clones Encoding Human PRO768

[2211] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43448. Based on the DNA43448consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO768.

[2212] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-GGCTGACACCGCAGTGCTCTTCAG-3′; (SEQ ID NO:438)reverse PCR primer 5′-GCTGCTGGGGACTGCAATGTAGCTG-3′. (SEQ ID NO:439)

[2213] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA43448 consensus sequence which had the followingnucleotide sequence:

[2214] hybridization probe

[2215] 5′-CATCCTCCATGTCTCCCATGAGGTCTCTAITGCrCCACGAAGCATC-3′ (SEQ IDNO:440).

[2216] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO768 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrowtissue (LIB255).

[2217] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO768 [herein designated as UNQ406(DNA55737-1345)] (SEQ ID NO:436) and the derived protein sequence forPRO768.

[2218] The entire nucleotide sequence of UNQ406 (DNA55737-1345) is shownin FIG. 176 (SEQ ID NO:436). Clone UNQ406 (DNA55737-1345) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 20-22 and ending at the stop codon at nucleotidepositions 3443-3445 (FIG. 176). The predicted polypeptide precursor is1141 amino acids long (FIG. 177). The full-length PRO768 protein shownin FIG. 177 has an estimated molecular weight of about 124,671 daltonsand a pI of about 5.82. Clone UNQ406 (DNA55737-1345) has been depositedwith the ATCC on Apr. 6, 1998. Regarding the sequence, it is understoodthat the deposited clone contains the correct sequence, and thesequences provided herein are based on known sequencing techniques.

[2219] Analysis of the amino acid sequence of the full-length PRO768polypeptide suggests that portions of it possess significant sequenceidentity and similarity with integrin 7.

[2220] Still analyzing the amino acid sequence of SEQ ID NO:437, theputative signal peptide is at about amino acids 1-33 of SEQ ID NO:437.The transmembrane domain is at amino acids 1039-1064 of SEQ ID NO:437.N-glycosylation sites are at amino acids: 86-89, 746-749, 949-952,985-988 and 1005-1008 of SEQ ID NO:437. Integrin alpha chain proteindomains are identified at about amino acids: 1064-1071, 384-409,1041-1071, 317-346, 443-465, 385-407, 215-224, 634-647, 85-99, 322-346,470-479, 442-466, 379-408 and 1031-1047 of SEQ ID NO:437. Thecorresponding nucleotides can be routinely determined given thesequences provided herein.

Example 71 Isolation of cDNA Clones Encoding Human PRO771

[2221] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA43330. Based on the DNA43330sequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence for PRO771.

[2222] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CAGCAATATTCAGAAGCGGCAAGGG-3′; (SEQ ID NO:443)reverse PCR primer 5′-CATCATGGTCATCACCACCATCATCATC-3′. (SEQ ID NO:444)

[2223] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA43330 consensus sequence which had the followingnucleotide sequence:

[2224] hybridization probe

[2225] 5′-GGTTACTACAAGCCAACACAATGTCATGGCAGTGTTGGACAGTGCTGG-3 ′ (SEQ IDNO:445).

[2226] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO771 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB28).

[2227] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO771 [herein designated as UNQ409(DNA49829-1346)] (SEQ ID NO:441) and the derived protein sequence forPRO771.

[2228] The entire nucleotide sequence of UNQ409 (DNA49829-1346) is shownin FIG. 178 (SEQ ID NO:441). Clone UNQ409 (DNA49829-1346) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 134-136 and ending at the stop codon atnucleotide positions 1442-1444 (FIG. 178). The predicted polypeptideprecursor is 436 amino acids long (FIG. 179). The full-length PRO771protein shown in FIG. 179 has an estimated molecular weight of about49,429 daltons and a pI of about 4.8. Clone UNQ409 (DNA49829-1346) hasbeen deposited with the ATCC on Apr. 7, 1998. Regarding the sequence, itis understood that the deposited clone contains the correct sequence,and the sequences provided herein are based on known sequencingtechniques.

[2229] Analysis of the amino acid sequence of the full-length PRO771polypeptide suggests that portions of it possess significant homology tothe testican protein, thereby indicating that PRO771 may be a noveltestican homologue.

[2230] Still analyzing the amino acid sequence of SEQ ID NO:442, theputative signal peptide, leucine zipper pattern, N-myristoylation sites,and thyroglobulin type-1 repeats are also shown in FIG. 179. Thecorresponding nucleotides can be routinely determined given thesequences provided herein.

Example 72 Isolation of cDNA Clones Encoding Human PRO733

[2231] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above, wherein the consensussequence obtained is herein designated DNA45600. Based on the DNA45600consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO733.

[2232] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-CCCAGCAGGGATGGGCGACAAGA-3′; (SEQ ID NO:448)reverse PCR primer 5′-GTCTTCCAGTTTCATATCCAATA-3′. (SEQ ID NO:449)

[2233] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA45600 consensus sequence which had the followingnucleotide sequence:

[2234] hybridization probe

[2235] 5′-CCAGAAGGAGCACGGGGAAGGGCAGCCAGATCTTGTCGCCCAT-3′ (SEQ IDNO:450).

[2236] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO733 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrowtissue (LIB255).

[2237] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO733 [herein designated as UNQ411(DNA52196-1348)] (SEQ ID NO:446) and the derived protein sequence forPRO733.

[2238] The entire nucleotide sequence of UNQ411 (DNA52196-1348) is shownin FIG. 180 (SEQ ID NO:446). Clone UNQ141 (DNA52196-1348) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 106-108 and ending at the stop codon atnucleotide positions 793-795 (FIG. 180). The predicted polypeptideprecursor is 229 amino acids long (FIG. 181). The full-length PRO733protein shown in FIG. 181 has an estimated molecular weight of about26,017 daltons and a pI of about 4.73. Clone UNQ411 (DNA52196-1348) hasbeen deposited with the ATCC on Apr. 7, 1998. Regarding the sequence, itis understood that the deposited clone contains the correct sequence,and the sequences provided herein are based on known sequencingtechniques.

[2239] Analysis of the amino acid sequence of the full-length PRO733polypeptide suggests that portions of it possess significant sequenceidentity and similarity to the T1/ST2 receptor binding protein precursorand therefore may have a similar function in cell signaling. If it is acytokine, it may be useful in the treatment of inflammation and cancer.

[2240] Still analyzing the amino acid sequence of SEQ ID NO:447, theputative signal peptide, transmembrane domain, N-myristoylation site,and tyrosine kinase site are also shown in FIG. 181. The correspondingnucleotides can be routinely determined given the sequences providedherein.

Example 73 Isolation of cDNA Clones Encoding Human PRO162

[2241] An expressed sequence tag (EST) DNA database (Merck/WashingtonUniversity) was searched and an EST AA397543 was identified which showedhomology to human pancreatitis-associated protein. The EST AA397543 colewas purchased and its insert obtained and sequenced and the sequenceobtained is shown in FIG. 182 (SEQ ID NO:451).

[2242] The entire nucleotide sequence of PRO162 is shown in FIG. 182(SEQ ID NO:451). DNA sequencing of the clone gave the full-length DNAsequence for PRO162 [herein designated as UNQ429 (DNA56965-1356)] (SEQID NO:451) and the derived protein sequence for PRO162. Clone UNQ429(DNA56965-1356) contains a single open reading frame with an apparenttranslational initiation site at nucleotide positions 86-88 and endingat the stop codon at nucleotide positions 611-613 (FIG. 182). Thepredicted polypeptide precursor is 175 amino acids long (FIG. 183). Thefull-length PRO162 protein shown in FIG. 183 has an estimated molecularweight of about 19,330 daltons and a pI of about 7.25. Clone UNQ429(DNA56965-1356) has been deposited with the ATCC. Regarding thesequence, it is understood that the deposited clone contains the correctsequence, and the sequences provided herein are based on knownsequencing techniques.

[2243] Analysis of the amino acid sequence of the full-length PRO162polypeptide suggests that portions of it possess significant homology tothe human pancreatitis-associated protein, thereby indicating thatPRO162 may be a novel pancreatitis-associated protein.

[2244] Still analyzing the amino acid sequence of SEQ ID NO:452, theputative signal peptide is at about amino acids 1-26 of SEQ ID NO:452. AC-type lectin domain signature is at about amino acids 146-171 of SEQ IDNO:452. The corresponding nucleotides can be routinely determined giventhe sequences provided herein.

Example 74 Isolation of cDNA Clones Encoding Human PRO788

[2245] A consensus DNA sequence (designated herein as DNA49308) wasassembled relative to other EST sequences using phrap as described inExample 1 above. Based upon an observed homology between the DNA49308consensus sequence and the Incyte EST clone no. 2777282, the Incyte ESTclone no. 2777282 was purchased and its insert obtained and sequenced,which gave the full-length DNA sequence for PRO788 [herein designated asUNQ430 (DNA56405-1357)] (SEQ ID NO:453) and the derived protein sequencefor PRO788.

[2246] Clone UNQ430 (DNA56405-1357) contains a single open reading framewith an apparent translational initiation site at nucleotide positions84-86 and ending at the stop codon at nucleotide positions 459-461 (FIG.184). The predicted polypeptide precursor is 125 amino acids long (FIG.185). The full-length PRO788 protein shown in FIG. 185 has an estimatedmolecular weight of about 13,115 daltons and a pI of about 5.90. CloneUNQ430 (DNA56405-1357) has been deposited with the ATCC. Regarding thesequence, it is understood that the deposited clone contains the correctsequence, and the sequences provided herein are based on knownsequencing techniques.

[2247] Still analyzing FIG. 185, a signal peptide is shown at aboutamino acids 1-17 of SEQ ID NO:454. An N-glycosylation site is at aboutamino acids 46-49 of SEQ ID NO:454.

Example 75 Isolation of cDNA Clones Encoding Human PRO1008

[2248] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated as DNA49804. An EST proprietary toGenentech was employed in the consensus assembly and is hereindesignated as DNA16508 (FIG. 188; SEQ ID NO:457). Based upon an observedhomology between the DNA49804 sequence and Merck EST clone no. AA143670,the Merck EST clone no. AA143670 was purchased and its insert obtainedand sequenced. That sequence is shown herein in FIG. 186 (SEQ IDNO:455).

[2249] Sequencing gave the full length sequence for PRO1008 [hereindesignated as UNQ492 (DNA57530-1375)] (SEQ ID NO:455) and the derivedprotein sequence for PRO1008 were identified.

[2250] The entire nucleotide sequence of UNQ492 (DNA57530-1375) is shownin FIG. 186 (SEQ ID NO:455). Clone UNQ492 (DNA57530-1375) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 138-140 and ending at the stop codon atnucleotide positions 936-938 (FIG. 186). The predicted polypeptideprecursor is 266 amino acids long (FIG. 187). The full-length PRO1008protein shown in FIG. 187 has an estimated molecular weight of about28,672 daltons and a pI of about 8.85. Clone UNQ492 (DNA57530-1375) hasbeen deposited with the ATCC on May 20, 1998. Regarding the sequence, itis understood that the deposited clone contains the correct sequence,and the sequences provided herein are based on known sequencingtechniques.

[2251] Analysis of the amino acid sequence of the full-length PRO1008polypeptide suggests that portions of it possess significant sequenceidentity and/or similarity with mdkk-1, thereby indicating that PRO1008may be a novel member of this family and have head inducing activity.

[2252] Still analyzing the amino acid sequence of SEQ ID NO:456, theputative signal peptide is at about amino acids 1-23 of SEQ ID NO:456.The N-glycosylation site is at about amino acids 256-259 of SEQ IDNO:456, and the fungal zn-(2)-cys(6) binuclear cluster domain is atabout amino acids 110-126 of SEQ ID NO:456. The correspondingnucleotides can of all the amino acids can be routinely determined giventhe sequences provided herein.

Example 76 Isolation of cDNA Clones Encoding Human PRO1012

[2253] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above, wherein theconsensus sequence is herein designated DNA49313. Based on the DNA49313consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO1012.

[2254] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-ACTCCCCAGGCTGTTCACACTGCC-3′ (SEQ ID NO:460);reverse PCR primer 5′-GATCAGCCAGCCAATACCAGCAGC-3′ (SEQ ID NO:461).

[2255] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA49313 consensus sequence which had the followingnucleotide sequence:

[2256] hybridization probe

[2257] 5′-GTGGTGATGATAGAATGCTTTGCCGAATGAAAGGAGTCAACAGCTATCCC-3′ (SEQ IDNO:462).

[2258] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1012 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[2259] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1012 [herein designated as UNQ495(DNA56439-1376)] (SEQ ID NO:458) and the derived protein sequence forPRO1012.

[2260] The entire nucleotide sequence of UNQ495 (DNA56439-1376) is shownin FIG. 189 (SEQ ID NO:458). Clone UNQ495 (DNA56439-1376) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 404-406 and ending at the stop codon atnucleotide positions 2645-2647 (FIG. 189). The predicted polypeptideprecursor is 747 amino acids long (FIG. 190). The full-length PRO1012protein shown in FIG. 190 has an estimated molecular weight of about86,127 daltons and apI of about 7.46. Clone UNQ495 (DNA56439-1376) hasbeen deposited with ATCC on May 14, 1998. Regarding the sequence, it isunderstood that the deposited clone contains the correct sequence, andthe sequences provided herein are based on known sequencing techniques.

[2261] Analysis of the amino acid sequence of the full-length PRO1012polypeptide suggests that portions of it possess sequence identity withdisulfide isomerase thereby indicating that PRO1012 may be a noveldisulfide isomerase related protein.

[2262] Still analyzing the amino acid sequence of SEQ ID NO:459, thecytochrome C family heme-binding site signature is at about amino acids158-163 of SEQ ID NO:459. The Nt-DNAJ domain signature is at about aminoacids 77-96 of SEQ ID NO:459. An N-glycosylation site is at about aminoacids 484-487 of SEQ ID NO:459. The ER targeting sequence is at aboutamino acids 744-747 of SEQ ID NO:459. It is understood that thepolypeptide and nucleic acids disclosed can be routinely formed with orwithout, these portions as desired, in alternative embodiments. Forexample, it may be desirable to produce PRO1012 without the ER targetingsequence. The corresponding nucleotides can be routinely determinedgiven the sequences provided herein.

Example 77 Isolation of cDNA Clones Encoding Human PRO1014

[2263] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above, wherein theconsensus sequence obtained is herein designated DNA49811. Based upon anobserved homology between the DNA49811 sequence and Incyte EST clone no.2612207, Incyte EST clone no. 2612207 was purchased and its insert wasobtained and sequenced, wherein the sequence obtained is shown in FIG.191 (SEQ ID NO:463).

[2264] DNA sequencing gave the full-length DNA sequence for PRO1014[herein designated as UNQ497 (DNA56409-1377)] (SEQ ID NO:463) and thederived protein sequence for PRO1014.

[2265] The entire nucleotide sequence of UNQ497 (DNA56409-1377) is shownin FIG. 191 (SEQ ID NO:463). Clone UNQ497 (DNA56409-1377) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 66-68 and ending at the stop codon at nucleotidepositions 966-968 (FIG. 191). The predicted polypeptide precursor is 300amino acids long (FIG. 192). The full-length PRO1014 protein shown inFIG. 192 has an estimated molecular weight of about 33,655 daltons and apI of about 9.31. Clone UNQ497 (DNA56409-1377) has been deposited withthe ATCC on May 20, 1998. Regarding the sequence, it is understood thatthe deposited clone contains the correct sequence, and the sequencesprovided herein are based on known sequencing techniques.

[2266] Analysis of the amino acid sequence of the full-length PRO1014polypeptide suggests that portions of it possess sequence identity withreductase, thereby indicating that PRO1014 may be a novel member of thereductase family.

[2267] Still analyzing the amino acid sequence of SEQ ID NO:464, theputative signal peptide is at about amino acids 1-19 of SEQ ID NO:464.The cAMP and cGMP dependent protein kinase phosphorylation sites are atabout amino acids 30-33 and 58-61 of SEQ ID NO:464. Short chain alcoholdehydrogenase family proteins are at about amino acids 165-202, 37-49,112-122 and 210-219 of SEQ ID NO:464. The corresponding nucleotides ofthese domains and any other amino acids provided herein can be routinelydetermined given the sequences provided herein.

Example 78 Isolation of cDNA Clones Encoding Human PRO1017

[2268] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above, wherein thatconsensus DNA sequence is herein designated DNA53235. Based upon anobserved homology between the DNA53235 consensus sequence and the MerckEST clone no. AA243086, the Merck EST clone no. AA243086 was purchasedand its insert obtained and sequenced, wherein the sequence obtained isshown in FIG. 193 (SEQ ID NO:465). DNA sequencing gave the full-lengthDNA sequence for PRO1017 [herein designated as UNQ500 (DNA56112-1379)](SEQ ID NO:465) and the derived protein sequence for PRO1017.

[2269] The entire nucleotide sequence of UNQ500 (DNA56112-1379) is shownin FIG. 193 (SEQ ID NO:465). Clone UNQ500 (DNA56112-1379) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 128-130 and ending at the stop codon atnucleotide positions 1370-1372 (FIG. 193). The predicted polypeptideprecursor is 414 amino acids long (FIG. 194). The full-length PRO1017protein shown in FIG. 194 has an estimated molecular weight of about48,414 daltons and a pI of about 9.54. Clone UNQ500 (DNA56112-1379) hasbeen deposited with the ATCC. Regarding the sequence, it is understoodthat the deposited clone contains the correct sequence, and thesequences provided herein are based on known sequencing techniques.

[2270] Analysis of the amino acid sequence of the full-length PRO1017polypeptide suggests that portions of it possess sequence identity withHNK-1 sulfotransferase, thereby indicating that PRO1017 may be a novelsulfotransferase.

[2271] Still analyzing the amino acid sequence of SEQ ID NO:466, theputative signal peptide is at about amino acids 1-31 of SEQ ID NO:466.N-glycosylation sites are at about amino acids 134-137, 209-212, 280-283and 370-273 of SEQ ID NO:466. The TNFR/NGFR family cystein-rich regionprotein is at about amino acids 329-332 of SEQ ID NO:466. Thecorresponding nucleotides can be routinely determined given thesequences provided herein. The protein can be secreted.

Example 79 Isolation of cDNA Clones Encoding Human PRO474

[2272] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above, wherein theconsensus sequence obtained is herein designated DNA49818. Based upon anobserved homology between the DNA49818 consensus sequence and the MerckEST clone no. H77889, the Merck EST clone no. H77889 was purchased andits insert obtained and sequenced, wherein the sequence obtained isherein shown in FIG. 195 (SEQ ID NO:467). DNA sequencing gave thefull-length DNA sequence for PRO474 [herein designated as UNQ502(DNA56045-1380)] (SEQ ID NO:467) and the derived protein sequence forPRO474.

[2273] The entire nucleotide sequence of UNQ502 (DNA56045-1380) is shownin FIG. 195 (SEQ ID NO:467). Clone UNQ502 (DNA56045-1380) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 106-108 and ending at the stop codon atnucleotide positions 916-918 (FIG. 195). The predicted polypeptideprecursor is 270 amino acids long (FIG. 196). The full-length PRO474protein shown in FIG. 196 has an estimated molecular weight of about28,317 daltons and a pI of about 6.0. Clone UNQ502 (DNA56045-1380) hasbeen deposited with the ATCC. Regarding the sequence, it is understoodthat the deposited clone contains the correct sequence, and thesequences provided herein are based on known sequencing techniques.

[2274] Still analyzing the amino acid sequence of SEQ ID NO:468, anN-glycosylation site is at about amino acids 138-141 of SEQ ID NO:468.Short-chain alcohol dehydrogenase family proteins are at about aminoacids 10-22, 81-91, 134-171 and 176-185 of SEQ ID NO:468. Thecorresponding nucleotides can be routinely determined given thesequences provided herein.

Example 80 Isolation of cDNA Clones Encoding Human PRO1031

[2275] An initial consensus DNA sequence was assembled relative to otherEST sequences using phrap as described in Example 1 above, wherein theconsensus sequence obtained is herein designated as DNA47332. Based uponan observed homology between the DNA47332 sequence and the Merck ESTclone no. W74558, Merck EST clone no. W74558 was purchased and itsinsert obtained and sequenced, wherein the sequence obtained is shown inFIG. 197 (SEQ ID NO:469). DNA sequencing gave the full-length DNAsequence for PRO1031 [herein designated as UNQ516 (DNA59294-1381)] (SEQID NO:469) and the derived protein sequence for PRO1031.

[2276] The entire nucleotide sequence of UNQ516 (DNA59294-1381) is shownin FIG. 197 (SEQ ID NO:469). Clone UNQ516 DNA59294-1381) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 4244 and ending at the stop codon at nucleotidepositions 582-584 (FIG. 197). The predicted polypeptide precursor is 180amino acids long (FIG. 198). The full-length PRO1031 protein shown inFIG. 198 has an estimated molecular weight of about 20,437 daltons and apI of about 9.58. Clone UNQ516 (DNA59294-1381) has been deposited withthe ATCC. Regarding the sequence, it is understood that the depositedclone contains the correct sequence, and the sequences provided hereinare based on known sequencing techniques.

[2277] Analysis of the amino acid sequence of the full-length PRO1031polypeptide suggests that it is a novel cytokine.

[2278] Still analyzing the amino acid sequence of SEQ ID NO:470, theputative signal peptide is at about amino acids 1-20 of SEQ ID NO:470.An N-glycosylation site is at about amino acids 75-78 of SEQ ID NO:470.A region having sequence identity with IL-17 is at about amino acids96-180. The corresponding nucleotides can be routinely determined giventhe sequences provided herein.

Example 81 Isolation of cDNA Clones Encoding Human PRO938

[2279] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above, wherein thatconsensus sequence is herein designated DNA49798. Based on the DNA49798DNA consensus sequence, oligonucleotides were synthesized: 1) toidentify by PCR a cDNA library that contained the sequence of interest,and 2) for use as probes to isolate a clone of the full-length codingsequence for PRO938.

[2280] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-GTCCAGCCCATGACCGCCTCCAAC-3′ (SEQ ID NO:473)reverse PCR primer 5′-CTCTCCTCATCCACACCAGCAGCC-3′ (SEQ ID NO:474)

[2281] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA49798 sequence which had the followingnucleotide sequence:

[2282] hybridization probe

[2283] 5′-GTGGATGCTGAAATTTACGCCCCATGGTGTCCATCCTGCCAGC-3′ (SEQ ID NO:475)

[2284] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO938 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue (LIB227).

[2285] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO938 [herein designated as UNQ475(DNA56433-1406)] (SEQ ID NO:471) and the derived protein sequence forPRO938.

[2286] The entire nucleotide sequence of UNQ475 (DNA56433-1406) is shownin FIG. 199 (SEQ ID NO:471). Clone UNQ475 (DNA56433-1406) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 134-136 and ending at the stop codon atnucleotide positions 1181-1183 (FIG. 199). The predicted polypeptideprecursor is 349 amino acids long (FIG. 200). The full-length PRO938protein shown in FIG. 200 has an estimated molecular weight of about38,952 daltons and a pI of about 4.34. Analysis of the full-lengthPRO938 sequence shown in FIG. 200 (SEQ ID NO:472) evidences the presenceof the following features: a signal peptide from amino 1 to about aminoacid 22, a transmembrane domain from about amino acid 191 to about aminoacid 211, a potential N-glycosylation site from about amino acid 46 toabout amino acid 49, a region homologous to disulfide isomerase fromabout amino acid 56 to about amino acid 72, and a region having sequenceidentity with flavodoxin proteins from about amino acid 173 to aboutamino acid 187.

[2287] Clone UNQ475 (DNA56433-1406) has been deposited with ATCC on May15, 1998, and is assigned ATCC Accession No. 209857.

[2288] Analysis of the amino acid sequence of the full-length PRO938polypeptide suggests that it possesses significant sequence similarityto protein disulfide isomerase, thereby indicating that PRO938 may be anovel protein disulfide isomerase. An analysis of the Dayhoff database(version 35.45 SwissProt 35) evidenced significant homology between thePRO938 amino acid sequence and the following Dayhoff sequences,P_W03626, P_WO3627, P_R70491, GARP_PLAFF, XLU85970_(—)1,ACADISPROA_(—)1, IE68_HSVSA, KSU52064_(—)1, U93872_(—)83, P_R97866.

Example 82 Isolation of cDNA Clones Encoding Human PRO1082

[2289] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above, wheein theconsensus sequence is herein designated DNA38097. Based on thisconsensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO1082.

[2290] A set of PCR primers (two forward and one reverse) weresynthesized: forward primer 1 5′-GTCCACAGACAGTCATCTCAGGAGCAG-3′ (SEQ IDNO:478); forward primer 2 5′-ACAAGTGTCTTCCCAACCTG-3′ (SEQ ID NO:479);reverse primer 1 5′-ATCCTCCCAGAGCCATGGTACCTC-3′ (SEQ ID NO:480).

[2291] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the DNA38097 consensus sequence which had the followingnucleotide sequence:

[2292] hybridization probe

[2293] 5 ′-CCAAGGATAGCTGTTGTTTCAGAGAAAGGATCGTGTGCTGCATCTCCTCCT-3′ (SEQID NO:481).

[2294] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primers identified above. A positive librarywas then used to isolate clones encoding the PRO1082 gene using theprobe oligonucleotide and one of the PCR primers. RNA for constructionof the cDNA libraries was isolated from human fetal kidney tissue(LIB227).

[2295] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1082 [herein designated as UNQ539(DNA53912-1457)] (SEQ ID NO:476) and the derived protein sequence forPRO1082.

[2296] The entire nucleotide sequence of UNQ539 (DNA53912-1457) is shownin FIG. 201 (SEQ ID NO:476). Clone UNQ539 (DNA53912-1457) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 160-162 and ending at the stop codon atnucleotide positions 763-765 (FIG. 201). The predicted polypeptideprecursor is 201 amino acids long (FIG. 202). The full-length PRO1082protein shown in FIG. 202 has an estimated molecular weight of about22,563 daltons and a pI of about 4.87. Clone UNQ539 (DNA53912-1457) hasbeen deposited with the ATCC. Regarding the sequence, it is understoodthat the deposited clone contains the correct sequence, and thesequences provided herein are based on known sequencing techniques.

[2297] Still analyzing the amino acid sequence of SEQ ID NO:477, thetransmembrane domain is at about amino acids 45-65 of SEQ ID NO:477. AcAMP- and cGMP-dependent protein kinase phosphorylation site is at aboutamino acids 197-200 of SEQ ID NO:477. N-myristoylation sites are atabout amino acids 3540 and 151-156 of SEQ ID NO:477. The regions whichshare sequence identity with the LDL receptor are at about amino acids34-67 and 70-200 of SEQ ID NO:477. The corresponding nucleotides ofthese amino acid regions and others can be routinely determined giventhe sequences provided herein.

Example 83 Isolation of cDNA Clones Encoding Human PRO1083

[2298] A cDNA sequence was identified using the amylase screeningtechnique described in Example 2 above, wherein that cDNA sequence isdesignated herein as DNA24256 (FIG. 205; SEQ ID NO:484). That cDNAsequence was then compared and aligned with other known EST sequences asdescribed in Example 1 above to obtain a consensus DNA sequence which isdesignated herein as DNA43422. Based on the DNA 43422 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO1083.

[2299] A pair of PCR primers (forward and reverse) were synthesized:forward PCR primer 5′-GGCATTGGAGCAGTGCTGGGTG-3′ (SEQ ID NO:485); reversePCR primer 5′-TGGAGGCCTAGATGCGGCTGGACG-3′ (SEQ ID NO:486).

[2300] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1083 gene usingthe reverse PCR primer. RNA for construction of the cDNA libraries wasisolated from human fetal kidney tissue (LIB227).

[2301] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1083 [herein designated as UNQ540(DNA50921-1458)] (SEQ ID NO:482) and the derived protein sequence forPRO1083.

[2302] The entire nucleotide sequence of UNQ540 (DNA50921-1458) is shownin FIG. 203 (SEQ ID NO:482). Clone UNQ540 (DNA50921-1458) contains asingle open reading frame with an apparent translational initiation siteat nucleotide positions 214-216 and ending at the stop codon atnucleotide positions 2293-2295 (FIG. 203). The predicted polypeptideprecursor is 693 amino acids long (FIG. 204). The full-length PRO1083protein shown in FIG. 204 has an estimated molecular weight of about77,738 daltons and a pI of about 8.87. Clone UNQ540 (DNA50921-1458) hasbeen deposited with the ATCC. Regarding the sequence, it is understoodthat the deposited clone contains the correct sequence, and thesequences provided herein are based on known sequencing techniques.

[2303] Still analyzing the amino acid sequence of SEQ ID NO:483, theputative signal peptide is at about amino acids 1-25 of SEQ I) NO:483.The transmembrane domains are at about amino acids 382-398, 402-420,445-468, 473-491, 519-537, 568-590 and 634-657 of SEQ ID NO:483. Amicrobodies C-terminal targeting signal is at about amino acids 691-693of SEQ ID NO:483. cAMP- and cGMP-dependent protein kinasephosphorylation sites are at about amino acids 198-201 and 370-373 ofSEQ ID NO:483. N-glycosylation sites are at about amino acids 39-42,148-151, 171-174, 234-237, 303-306, 324-227 and 341-344 of SEQ IDNO:483.

[2304] A G-protein coupled receptor family domain is at about aminoacids 475-504 of SEQ ID NO:483. The corresponding nucleotides can beroutinely determined given the sequences provided herein.

Example 84 Isolation of cDNA Clones Encoding Human PRO200

[2305] Probes based on an expressed sequence tag (EST) identified fromthe Incyte Pharmaceuticals database due to homology with VEGF were usedto screen a cDNA library derived from the human glioma cell line G61. Inparticular, Incyte Clone “INC1302516” was used to generate the followingfour probes: (SEQ ID ACTTCTCAGTGTCCATAAGGG; NO:489) (SEQ IDGAACTAAAGAGAACCGATACCATTTTCTGGCCAGGTTGTC; NO:490) (SEQ IDCACCACAGCGTTTAACCAGG; and NO:491) (SEQ ID ACAACAGGCACAGTTCCCAC NO:492).

[2306] Nine positives were identified and characterized. Three clonescontained the fill coding region and were identical in sequence. Partialclones were also identified from a fetal lung library and were identicalwith the glioma-derived sequence with the exception of one nucleotidechange which did not alter the encoded amino acid.

Example 85 Expression Constructs for PRO200

[2307] For mammalian protein expression, the entire open reading frame(ORF) was cloned into a CMV-based expression vector. An epitope-tag(FLAG, Kodak) and Histidine-tag (His8) were inserted between the ORF andstop codon. VEGF-E-His8 and VEGF-E-FLAG were transfected into humanembryonic kidney 293 cells by SuperFect (Qiagen) and pulse-labeled for 3hours with [³⁵S]methionine and [³⁵C]cysteine. Both epitope-taggedproteins co-migrate when 20 microliters of 15-fold concentratedserum-free conditioned medium were electrophoresed on a polyacrylamidegel (Novex) in sodium dodecyl sulfate sample buffer (SDS-PAGE). TheVEGF-E-IgG expression plasmid was constructed by cloning the ORF infront of the human Fc (IgG) sequence.

[2308] The VEGF-E-IgG plasmid was co-transfected with BaculogoldBaculovirus DNA (Pharmingen) using Lipofectin (GibcoBRL) into 10⁵ Sf9cells grown in Hink's TNM-FH medium (JRH Biosciences) supplemented with10% fetal bovine serum. Cells were incubated for 5 days at 28° C. Thesupernatant was harvested and subsequently used for the first viralamplification by infecting Sf9 cells at an approximate multiplicity ofinfection (MOI) of 10. Cells were incubated for 3 days, then supernatantharvested, and expression of the recombinant plasmid determined bybinding of 1 ml of supernatant to 30 μl of Protein-A Sepharose CL-4Bbeads (Pharmacia) followed by subsequent SDS-PAGE analysis. The firstamplification supernatant was used to infect a 500 ml spinner culture ofSf9 cells grown in ESF-921 medium (Expression Systems LLC) at anapproximate MOI of 0.1. Cells were treated as above, except harvestedsupernatant was sterile filtered. Specific protein was purified bybinding to Protein-A Sepharose 4 Fast Flow (Pharmacia) column.

Example 86 Northern Blot Analyses for PRO200

[2309] Blots of human poly(A)+ RNA from multiple adult and fetal tissuesand tumor cell lines were obtained from Clontech (Palo Alto, Calif.).Hybridization was carried out using ³²P-labeled probes containing theentire coding region and washed in 0.1× SSC, 0.1% SDS at 63° C.

[2310] VEGF-E mRNA was detectable in fetal lung, kidney, brain, liverand adult heart, placenta, liver, skeletal muscle, kidney, and pancreas.VEGF-E mRNA was also found in A549 lung adenocarcinoma and HeLa cervicaladenocarcinoma cell lines.

Example 87 In Situ Hybridization of Human Fetal Tissue Sections forPRO200

[2311] Formalin-fixed, paraffin-embedded human fetal brain, liver, lowerlimb, small intestine, thyroid, lymph node, thymus, stomach, trachea,skin, spleen, spinal cord, adrenal, placenta, cord, and adult liver,pancreas, lung, spleen, lymph node, adrenal, heart, aorta, and skin weresectioned, deparaffinized, deproteinated in proteinase K (20 μg/ml) for15 minutes at 37° C., and further processed for in situ hybridization asdescribed by Lu LH and Gillett NA (Cell Vision 1:169-176, 1994). A[α³³-P]UTP-labeled antisense riboprobe was generated from a PCR productof 980 bp (primers GGCGGAATCCAACCTGAGTAG and GCGGCTATCCTCCTGTGCTC, SEQID NOS:493 and494, respectively). The slides were dipped in Kodak NTB2nuclear track emulsion and exposed for 4 weeks.

[2312] VEGF-E mRNA expression included localization at the growth plateregion and embracing fetal myocytes.

Example 88 Myocyte Hyertrophy Assay for PRO200

[2313] Myocytes from neonatal Harlan Sprague Dawley rat heart ventricle(23 days gestation) were plated in duplicate at 75000 cells/ml in a96-well plate. Cells were treated for 48h with 2000, 200, 20, or 2 ng/mlVEGF-E-IgG. Myocytes were stained with crystal violet to visualizemorphology and scored on a scale of 3 to 7, 3 being nonstimulated and 7being full-blown hypertrophy.

[2314] 2000 ng/ml and 200 ng/ml VEGF-E caused hypertrophy, scored as a5.

Example 89 Cell Proliferation Assav for PRO200

[2315] Mouse embryonic fibroblast C3HIOT1/2 cells (ATCC) were grown in50:50 Ham's F-12: low glucose DMEM medium containing 10% fetal calfserum (FCS). Cells were plated in duplicate in a 24-well plate at 1000,2000, and 4000 cells/well. After 48 hours, cells were switched to mediumcontaining 2% FCS and were incubated for 72 hours with 200, 800, or 2000ng/ml VEGF-E or no growth factor added.

[2316] Approximately 1.5 fold greater number of cells were measured inthe presence of 200 ng/ml VEGF-E as in its absence, at all three celldensities.

Example 90 Endothelial Cell Survival Assay for PRO200

[2317] Human umbilical vein endothelial cells (HUVEC, Cell Systems) weremaintained in Complete Media (Cell Systems) and plated in triplicate inserum-free medium (Basic Media from Cell Systems containing 0.1% BSA) at20,000 cells/well of a 48-well plate. Cells were incubated for 5 dayswith 200 or 400 ng/ml VEGF-E-IgG, 100 ng/ml VEGF, 20 ng/ml basic FGF, orno addition.

[2318] Survival was 2-3 times greater with VEGF-E as compared to lack ofgrowth factor addition. VEGF and basic FGF were included as positivecontrols.

Example 91 Isolation of cDNA Clones Encoding Human PRO285

[2319] A proprietary expressed sequence tag (EST) DNA database LIFESEQ™,Incyte Pharmaceuticals, Palo Alto, Calif.) was searched and an EST(#2243209) was identified which showed homology to the Drosophila Tollprotein.

[2320] Based on the EST, a pair of PCR primers (forward and reverse):TAAAGACCCAGCTGTGACCG (SEQ ID NO:499) ATCCATGAGCCTCTGATGGG (SEQ IDNO:500), and a probe: ATTTATGTCTCGAGGAAAGGGACTGGTTACCAGGGCAGCCAGTTC (SEQID NO:501)

[2321] were synthesized.

[2322] mRNA for construction of the cDNA libraries was isolated fromhuman placenta tissue. The cDNA libraries used to isolate the cDNAclones were constructed by standard methods using commercially availablereagents such as those from Invitrogen, San Diego, Calif. (Fast Track2). The cDNA was primed with oligo dT containing a NotI site, linkedwith blunt to SalI hemikinased adaptors, cleaved with NotI, sizedappropriately by gel electrophoresis, and cloned in a definedorientation into the cloning vector pCR2.1 (Invitrogen, Inc.) usingreagents and protocols from Life Technologies, Gaithersburg, Md. (SuperScript Plasmid System). The double stranded cDNA was sized to greaterthan 1000 bp and the cDNA was cloned into BamHI/NotI cleaved vector.pCR2.1 is a commercially available plasmid, designed for easy cloning ofPCR fragments, that carries AmpR and KanR genes for selection, and LacZgene for blue-white selection.

[2323] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO285 gene usingthe probe oligonucleotide and one of the PCR primers.

[2324] A cDNA clone was sequenced in entirety. The entire nucleotidesequence of DNA40021-1154 (encoding PRO285) is shown in FIG. 208 (SEQ IDNO:495). Clone DNA40021-1154 contains a single open reading frame withan apparent translational initiation site at nucleotide positions 61-63(FIG. 208). The predicted polypeptide precursor is 1049 amino acidslong, including a putative signal peptide at amino acid positions 1-29,a putative transmembrane domain between amino acid positions 837-860,and a leucine zipper pattern at amino acid positions 132-153 and704-725, respectively. It is noted that the indicated boundaries areapproximate, and the actual limits of the indicated regions might differby a few amino acids. Clone DNA40021-1154 has been deposited with ATCC(designation: DNA40021-1154) and is assigned ATCC deposit no.209389.

[2325] Based on a BLAST and FastA sequence alignment analysis (using theALIGN computer program) of the full-length sequence is a human analogueof the Drosophila Toll protein, and is homologous to the following humanToll proteins: Toll (DNAX# HSU88540-1, which is identical with therandom sequenced full-length cDNA #HUMRSC786-1); Tllo2 (DNAX#HSU88878-1); Toll3 (DNAX# HSU88879-1); and Toll4 (DNAX# HSU88880-1).

Example 92 Isolation of cDNA Clones Encodin Human PRO286

[2326] A proprietary expressed sequence tag (EST) DNA database(LIFESEQ™, Incyte Pharmaceuticals, Palo Alto, Calif.) was searched andan EST (#694401) was identified which showed homology to the DrosophilaToll protein.

[2327] Based on the EST, a pair of PCR primers (forward and reverse):GCCGAGACAAAAACGTTCTCC (SEQ ID NO:502) CATCCATGTTCTCATCCATTAGCC (SEQ IDNO:503), and a probe: TCGACAACCTCATGCAGAGCATCAACCAAAGCAAGAAAACAGTATT(SEQ ID NO: 504)

[2328] were synthesized.

[2329] mRNA for construction of the cDNA libraries was isolated fromhuman placenta tissue. This RNA was used to generate an oligo dT primedcDNA library in the vector pRK5D using reagents and protocols from LifeTechnologies, Gaithersburg, Md. (Super Script Plasmid System). pRK5D isa cloning vector that has an sp6 transcription initiation site followedby an SfiI restriction enzyme site preceding the XhoI/NotI cDNA cloningsites. The cDNA was primed with oligo dT containing a NotI site, finkedwith blunt to SalI hemikinased adaptors, cleaved with NotI, sized togreater than 1000 bp appropriately by gel electrophoresis, and cloned ina defined orientation into XhoI/NotI-cleaved pRK5D.

[2330] Ea order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO286 gene usingthe probe oligonucleotide identified above and one of the PCR primers.

[2331] A cDNA clone was sequenced in entirety. The entire nucleotidesequence of DNA42663-1154 (encoding PRO286) is shown in FIG. 210 (SEQ IDNO:497). Clone DNA42663-1154 contains a single open reading frame withan apparent translational initiation site at nucleotide positions 57-59(FIG. 211). The predicted polypeptide precursor is 1041 amino acidslong, including a putative signal peptide at amino acid positions 1-26,a potential transmembrane domain at amino acid positions 826-848, andleucine zipper patterns at amino acids 130-151, 206-227, 662-684,669-690 and 693-614, respectively. It is noted that the indicatedboundaries are approximate, and the actual limits of the indicatedregions might differ by a few amino acids. Clone DNA42663-1154 has beendeposited with ATCC (designation: DNA42663-1154) and is assigned ATCCdeposit no. 209386.

[2332] Based on a BLAST and FastA sequence alignment analysis (using theALIGN computer program) of the full-length sequence of PRO286, it is ahuman analogue of the Drosophila Toll protein, and is homologous to thefollowing human Toll proteins: Toll1 (DNAX# HSU88540-1, which isidentical with the random sequenced full-length cDNA #HUMRSC786-1);Toll2 (DNAX# HSU88878-1); To113 (DNAX# HSU88879-1); and Toll4 (DNAX#HSU88880-1).

Example 93 NF-κB Assay for PRO285 and PRO286

[2333] As the Toll proteins signal through the NF-κB pathway, theirbiological activity can be tested in an NF-κB assay. In this assayJurkat cells are transiently transfected using Lipofectamine reagent(Gibco BRL) according to the manufacturer's instructions. 1 μg pB2XLucplasmid, containing NF-κB driven luciferase gene, is contransfected with1 μg pSRαN expression vector with or without the insert encoding PRO285or PRO286. For a positive control, cels are treated with PMA (phorbolmyristyl acetate; 20 ng/ml) and PHA (phytohaemaglutinin, 2 μg/ml) forthree to four hours. Cells are lysed 2 or 3 days later for measurementof luciferase activity using reagents from Promega.

Example 94 Isolation of cDNA Clones Encoding Human PRO213-1. PRO1330 andPRO1449

[2334] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA28735. Based on the DNA28735 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO213-1, PRO1330 and/or PRO1449. A pair of PCR primers (forward andreverse) were synthesized: forward PCR primer5′-TGGAGCAGCAATATGCCAGCC-3′ (SEQ ID NO:511) reverse PCR primer5′-TTTTCCACTCCTGTCGGGTTGG-3′ (SEQ ID NO:512)

[2335] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA28735 sequence which had the followingnucleotide sequence:

[2336] hybridization probe

[2337] 5′-GGTGACACTTGCCAGTCAGATGTGGATGAATGCAGTGCTAGGAGGG-3′ (SEQ IDNO:513)

[2338] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO213-1, PRO1330and/or PRO1449 gene using the probe oligonucleotide and one of the PCRprimers. RNA for construction of the cDNA libraries was isolated fromhuman fetal lung tissue.

[2339] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence encoding PRO213-1, PRO1330 and/or PRO1449(DNA30943-1-1163-1 (SEQ ID NO:505), DNA64907-1163-1 (SEQ ID NO:507) andDNA64908-1163-1 (SEQ ID NO:509), respectively].

[2340] The entire nucleotide sequences corresponding toDNA30943-1-1163-1 (SEQ ID NO:505), DNA64907-1163-1 (SEQ ID NO:507) andDNA64908-1163-1 (SEQ ID NO:509), respectively. DNA30943-1163,DNA64907-1163-1 and DNA64908-1163-1 contain a single open reading framewith an apparent translational initiation site at nucleotide positions336-338, 488-490 and 326-328, respectively, and ending at the stop codonat nucleotide positions 1221-1223, 1307-1309 and 1145-1147, respectively(FIGS. 212, 214 and 216). The predicted polypeptide precursor is 295,273 and 273 amino acids long, respectively (FIGS. 213, 215 and 217).DNA30943-1-1163-1, DNA64907-1163-1 and DNA64908-1163-1 have beendeposited with ATCC and are assigned ATCC deposit no. 209791, 203242 and203243, respectively.

[2341] Analysis of the amino acid sequence of the full-length PRO213-1polypeptide suggests that a portion of it possess significant homologyto the human growth arrest-specific gene 6 protein. More specifically,an analysis of the Dayhoff database (version 35.45 SwissProt 35)evidenced significant homology between the PRO213 amino acid sequenceand the following Dayhoff sequences, HSMHC3W5A_(—)6 and B48089.

[2342] Additional analysis of the amino acid sequence of the full-lengthPRO1330 and PRO1449 polypeptide indicates significant identity withnotch4. More specifically, an analysis of the Dayhoff database (version35.130 SwissProt 35) evidenced significant identity between PRO1330 andthe following Dayhoff sequences, D86566_(—)1 and NEL_HUMAN.

Example 95 Isolation of cDNA Clones Encoding Human PRO298

[2343] A cDNA isolated in the amylase screen described in Example 2above is herein designated DNA26832 (FIG. 220; SEQ ID NO:516). Thesequence of DNA26832 was then used to search expressed sequence tag(EST) databases. The EST databases included public EST databases (e.g.,GenBank) and a proprietary EST database (LIFESEQ™, IncytePharmaceuticals, Palo Alto, Calif.). The search was performed using thecomputer program BLAST or BLAST2 (Altshul et al., Methods in Enzymology266: 469-480 [1996]). Those comparisons resulting in a BLAST score of 70(or in some cases 90) or greater that did not encode proteins wereclustered and assembled into consensus DNA sequences with the program“phrap” (Phil Green, University of Washington, Seattle, Wash.;http:/Ibozeman.mbt.washington.edu/phrap.docs/phrap.html).

[2344] A consensus DNA sequence was assembled relative to other ESTsequences using phrap. A consensus sequence was determined, which wasthen extended using repeated cycles of BLAST and phrap to extend theconsensus sequence as far as possible using the sources of EST sequencesdiscussed above. The extended assembly sequence was designatedDNA35861.Based on the DNA35861 consensus sequence, oligonucleotides weresynthesized: 1) to identity by PCR a cDNA library that contained thesequence of interest, and 2) for use as probes to isolate a clone of thefull-length coding sequence of PRO298. Forward and reverse primersgenerally range from 20 to 30 nucleotides and are often designed to givea PCR product of about 100-1000 bp in length. The probe sequence istypically 40-55 bp in length. In some cases, additional oligonucleotidesare synthesized when the consensus sequence is greater than about 1-1.5kbp. In order to screen several libraries for a full-length clone, DNAfrom the libraries was screened by PCR amplification, as per Ausubel etal., Current Protocols in Molecular Biology, with the PCR primer pair. Apositive library was used to isolate clones encoding the gene ofinterest using the probe oligonucleotide and one of the primer pairs.

[2345] PCR primers (forward and reverse) and a hybridization probe weresynthesized: forward PCR primer 1 CAACGTGATTTCAAAGCTGGGCTC (SEQ IDNO:517) forward PCR primer 2 GCCTCGTATCAAGAATTTCC (SEQ ID NO:518)forward PCR primer 3 AGTGGAAGTCGACCTCCC (SEQ ID NO:519) reverse PCRprimer 1 CTCACCTGAAATCTCTCATAGCCC (SEQ ID NO:520) hybridization probe 1CGCAAAACCCATTTTGGGAGCAGGAATTCCAATCATGTCTGTGATGGTGG (SEQ ID NO:521)

[2346] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO298 gene usingthe probe oligonucleotide and one of the PCR primers.

[2347] RNA for construction of the cDNA libraries was isolated fromhuman fetal lung tissue (LIB25). The cDNA libraries used to isolated thecDNA clones were constructed by standard methods using commerciallyavailable reagents such as those from Invitrogen, San Diego, Calif. ThecDNA was primed with oligo dT containing a Nod site, linked with bluntto Sail hemikinased adaptors, cleaved with NotI, sized appropriately bygel electrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the Sfil site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

[2348] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO298 (herein designated UNQ261(DNA39975-12101) (SEQ ID NO:514), and the derived protein sequence forPRO298 (SEQ ID NO:515).

[2349] The entire nucleotide sequence of UNQ261 (DNA39975-1210) is shownin FIG. 218 (SEQ ID NO:514). Clone DNA39975-1210 contains a single openreading frame with an apparent translational initiation site atnucleotide positions 375-377. The predicted polypeptide precursor is 364amino acids long. The protein contains four putative transmembranedomains between amino acid positions 36-55 (type II TM), 65-84, 188-208,and 229-245, respectively. A putative N-linked glycosylation site startsat amino acid position 253. In addition, the following features havebeen identified in the protein sequence: cAMP- and cGMP-dependentprotein kinase phosphorylation site, starting at position 8;N-myristoylation sites starting a position 173 and 262, respectively;and a ZP domain between amino acid positions 45-60. Clone DNA39975-1210has been deposited with ATCC (Apr. 21, 1998) and is assigned ATCCdeposit no.209783.

Example 96 Isolation of cDNA Clones Encoding Human PRO337

[2350] A cDNA sequence identified in the amylase screen described inExample 2 above is herein designated DNA42301 (FIG. 223, SEQ ID NO:524).The DNA42301 sequence was then compared to other EST sequences usingphrap as described in Example 1 above and a consensus sequencedesignated herein as DNA28761 was identified. Based on this consensussequence, oligonucleotides were synthesized: 1) to identify -by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence. In orderto screen several libraries for a source of a full-length clone, DNAfrom the libraries was screened by PCR amplification with the PCR primerpair identified above. A positive library was then used to isolateclones encoding the PRO337 gene using the probe oligonucleotide and oneof the PCR primers. RNA for construction of the cDNA libraries wasisolated from human fetal brain.

[2351] A cDNA clone was sequenced in its entirety. The full lengthnucleotide sequence of DNA43316-1237 is shown in FIG. 221 (SEQ IDNO:522). Clone DNA43316-1237 contains a single open reading frame withan apparent translational initiation site at nucleotide positions134-136 (FIG. 221; SEQ ID NO:522). The predicted polypeptide precursoris 344 amino acids long. Clone DNA43316-1237 has been deposited withATCC and is assigned ATCC deposit no. 209487.

[2352] Based on a BLAST-2 and FastA sequence alignment analysis of thefull-length sequence, PRO337 shows amino acid sequence identity to ratneurotimin (97%).

Example 97 Isolation of cDNA Clones Encoding Human PRO403

[2353] Introduction:

[2354] Human thrombopoietin (THPO) is a glycosylated hormone of 352amino acids consisting of two domains. The N-terminal domain, sharing50% similarity to erythropoietin, is responsible for the biologicalactivity. The C-terminal region is required for secretion. The gene forthrombopoietin (ThPO) maps to human chromosome 3q27-q8 where the sixexons of this gene span 7 kilobase base pairs of genomic DNA (Chang etal., Genomics 26: 636-7 (1995); Foster et al., Proc. Natl. Acad. Sci.USA 91: 13023-7 (1994); Gurney et al., Blood 85: 981-988 (1995). Inorder to determine whether there were any genes encoding THPO homologueslocated in close proximity to THPO, genomic DNA fragments from thisregion were identified and sequenced. Three P1 clones and one PAC clones(Genome Systems Inc., St. Louis, Mo.; cat. Nos. P1-2535 and PAC-6539)encompassing the THPO locus were isolated and a 140 kb region wassequenced using the ordered shotgun strategy (Chenet al., Genomics 17:651-656 (1993)), coupled with a PCR-based gap filling approach. Analysisreveals that the region is gene-rich with four additional genes locatedvery close to THPO: tumor necrosis factor- receptor type 1 associatedprotein 2 (TRAP2) and elongation initiation factor gamma (elF4( ),chloride channel 2 (CLCN2) and RNA polymerase II subunit hRPB17. Whileno THPO homolog was found in the region, four novel genes have beenpredicted by computer-assisted gene detection (GRAIL)(Xu et al., Gen.Engin. 16: 241-253 (1994), the presence of CpG islands (Cross, S. andBird, A., Curr. Opin. Genet. & Devel. 5: 109-314 (1995), and homology toknown genes (as detected by WU-BLAST2.0)(Altschul and Gish, MethodsEnzymol. 266: 460-480 (1996) (http://blast.wustl.edu/blast/README.html).

[2355] Procedures:

[2356] P1 and PAC Clones:

[2357] The initial human P1 clone was isolated from a genomic P1 library(Genome Systems Inc., St. Louis, MO; cat. no.: P1-2535) screened withPCR primers designed from the THPO genomic sequence (A. L. Gurney, etal., Blood 85: 981-88 (1995). PCR primers were designed from the endsequences derived from this P1 clone were then used to screen P1 and PAClibraries (Genome Systems, Cat. Nos.: P1-2535 & PAC-6539) to identifyoverlapping clones (PAC1, p1.t, and P1.u). The 3′-end sequence fromPAC.z was used to define the primers used for the screening of a humanBAC library (Genome Systems Inc., St. Louis, MO; Cat. No.: BDTW-4533A).

[2358] Ordered Shotgtun Strategy:

[2359] The Ordered Shotgun Strategy (OSS) (Chen et al., Genomics 17:651-656 (1993)) involves the mapping and sequencing of large genomic DNAclones with a hierarchical approach. The P1 or PAC clone was sonicatedand the fragments subcloned into lambda vector (ABluestar) (Novagen,Inc., Madison, Wis.; cat. no. 69242-3). The lambda subclone inserts wereisolated by long-range PCR (Barnes, W. Proc. Natl. Acad. Sci. USA 91:2216-2220 (1994) and the ends sequenced. The lambda-end sequences wereoverlapped to create a partial map of the original clone. Those lambdaclones with overlapping end-sequences were identified, the insetssubcloned into a plasmid vector (pUC18 or pUC19, Hoefer PharmaciaBiotech, Inc., San Francisco, Calif., Cat. Nos. 27-4949-01 and274951-01) and the ends of the plasmid subclones were sequenced andassembled to generate a contiguous sequence. This directed sequencingstrategy minimizes the redundancy required while allowing one to scanfor and concentrate on interesting regions.

[2360] In order to define better the THPO locus and to search for othergenes related to the hematopoietin family, five genomic clones wereisolated from this region by PCR screening of human PI and PAC libraries(Genome System, Inc., Cat. Nos.: P1-2535 and PAC-6539). _os The sizes ofthe genomic fragments are as follows: P1.t is 40 kb; P1.g is 70 kb; P1.uis 70 kb; PAC.z is 200 kb; and BAC.1 is 80 kb. Approximately 75% (140kb) of the 190 kb genomic DNA region was sequenced by the OrderedShotgun Strategy (OSS) (Chen et al., Genomics 17: 651-56 (1993), andassembled into contigs using AutoAssemblerTM (Applied Biosystems, PerkinElmer, Foster City, Calif., cat. no. 903227). The preliminary order ofthese contigs was determined by manual analysis. There were 47 contigsthe 140 kb region. A PCR-based approach to ordering the contigs andfilling in the gaps was employed. The following summarizes the numberand sizes of the gaps. The 50 kb of sequence unique to BAC.1 wassequenced by a total shotgun approach with a ten-fold redundancy. Sizeof gap number <50 bp 13  50-150 bp  7  150-300 bp  7  300-1000 bp 101000-5000 bp  7 >5000 bp  2 ((15,000 bp)

[2361] DNA Sequencing:

[2362] ABI DYE-primerTM chemistry (PE Applied Biosystems, Foster City,Calif.; Cat. No.: 402112) was used to end-sequence the lambda andplasmid subclones. ABI DYE-terminaterTM chemistry (PE AppliedBiosystems, Foster City, Calif., Cat. No: 403044) was used to sequencethe PCR products with their respective PCR primers. The sequences werecollected with an ABI377 instrument. For PCR products larger than 1 kb,walking primers were used. The sequences of contigs generated by the OSSstrategy in AutoAssemblerTM (PE Applied Biosystems, Foster City, Calif.;Cat. No: 903227) and the gap-filling sequencing trace files wereimported into SequencherTM (Gene Codes Corp., Ann Arbor, Mich.) foroverlapping and editing. The sequences generated by the total shotgunstrategy were assembled using Phred and Phrap and edited using Consed(http://chimera.biotech.washington.eduluwgc/projects.htm) and GFP(Genome Reconstruction Manager for Phrap), version 1.2(http://stork.cellb.bcm.tmc.edu/gfp/).

[2363] PCR-Based Gap Filling Strategy:

[2364] Primers were designed based on the 5′- and 3′-end sequenced ofeach contig, avoiding repetitive and low quality sequence regions. Allprimers were designed to be 19-24-mers with 50-70% G/C content. Oligoswere synthesized and gel-purified by standard methods.

[2365] Since the orientation and order of the contigs were unknown,permutations of the primers were used in the amplification reactions.Two PCR kits were used: first, XL PCR kit (Perkin Elmer, Norwalk, Conn.;Cat. No.: N8080205), with extension times of approximately 10 minutes;and second, the Taq polymerase PCR kit (Qiagen Inc., Valencia, Calif.;Cat. No.: 201223) was used under high stringency conditions if smearedor multiple products were observed with the XL PCR kit. The main PCRproduct from each successful reaction was extracted from a 0.9% lowmelting agarose gel and purified with the Geneclean DNA Purification kitprior to sequencing.

[2366] Analysis:

[2367] The identification and characterization of coding regions wascarried out as follows: First, repetitive sequences were masked usingRepeatMasker (A. F. A. Smit & P. Green, http://flp.genome.washington.eduRM/RM_details.html) which screens DNA sequences in FastA format againsta library of repetitive elements and returns a masked query sequence.Repeats not masked were identified by comparing the sequence to theGenBank database using WUBLAST2.0 [Altschul, S & Gish, W., MethodsEnzymol. 266: 460-480 (1996); http://blast.wustl.edu/blast/README.html]and were masked manually.

[2368] Next, known genes were revealed by comparing the genomic regionsagainst Genentech's protein database using the WUBLAST2.0 algorithm andthen annotated by aligning the genomic and cDNA sequences for each gene,respectively, using a Needleman-Wunch (Needleman and Wunsch, J. Mol.Biol. 48: 443-453 (1970) algorithm to find regions of local identitybetween sequences. The strategy results in detection of all exons of thefive known genes in the region, THPO, TRAP2, eIF4g, CLCN2 and hRPB17(see below). Known genes Map position eukaryotic translation initiationfactor 4 gamma 3q27-qter thrombopoietin 3q26-q27 chloride channel 23q26-qter TNF receptor associated protein 2 not previously mapped RNApolymerase II subunit hRPB17 not previously mapped

[2369] Finally, novel transcription units were predicted using a numberof approaches. CpG islands (S. Cross & Bird, A., Curr. Opin. Genet. Dev.5: 109-314 (1995) islands were used to define promoter regions and wereidentified as clusters of sites cleaved by enzymes recognizing GC-rich,6 or 8-mer palindromic sequences (NotI, NarI, BssHII, XhoI. CpG islandsare usually associated with promoter regions of genes. WUBLAST2.0analysis of short genomic regions (10-20 kb) versus GenBank revealedmatches to ESTs. The individual EST sequences (or where possible, theirsequence chromatogram files) were retrieved and assembled with Sequencerto provide a theoretical cDNA sequence (DNA36443). GRAIL2 (ApoCom Inc.,Knoxville, Tenn., command line version for the DEC alpha) was used topredict a novel exon. The five known genes in the region served asinternal controls for the success of the GRAIL algorithm.

[2370] Isolation:

[2371] A partial endothelin converting enzyme-2 (ECE-2) cDNA clone wasisolated by first splicing in silico the ECE-2 exons predicted in thegenomic sequence to generate a putative sequence (DNA36443). Anoligonucleotide probe: GAAGCAGTGCAGCCAGCAGTAGAGAGGCACCTGCTAAGA) (SEQ IDNO:530) was designed and used to screen a human fetal small intestinelibrary (LIB110) and internal PCR primers (36443fl)(ECE2.f:ACGCAGCTGGAGCTGGTCTTAGCA) (SEQ ID NO:531) and (36443r1) (ECE2.r)(GGTACTGGACCCCTAGGGCCACAA) (SEQ ID NO:532) were used to confirm cloneshybridizing to the probe prior to sequencing. One positive clone wasobtained, however this cDNA (DNA49830) represented a partially splicedtranscript containing appropriately spliced exons 1 through 6, followedby intron 6 sequence. The oligo dT primer annealed to a polyA-stretchwithin an Alu element present in intron 6. An additional ECE-2 cDNAfragment (DNA49831) was obtained by PCR from a human fetal kidneylibrary (LIB227) with primers designed from the presumed cDNA sequence[36443f3: CCTCCCAGCCGAGACCAGTGG (SEQ ID NO:533) and 36443r2:GGTCCTATAAGGGCCAAGACC (SEQ ID NO:534)]. This PCR product extended fromexon 13 into the 3′ untranslated region in exon 18.

[2372] A full length endothelin converting enzyme 2 (ECE-2) cDNA clone(DNA55800-1263) was isolated from an oligo-dT-primed human fetal brainlibrary. RNA from human fetal brain tissue (20 weeks gestation,#283005)(SRC175) was isolated by guanidine thiocyanate and 5 μg used togenerate double stranded cDNA which was cloned into the vector pRK5E.The 3′-primer (pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTTT) (SEQ IDNO:535) and the 5-linker (pCGGACGCGTGGGTCGA) (SEQ ID NO:536) weredesigned to introduce XhoI and NotI restriction sites. The library wasscreened with PCR primers [36443pcrf1: CGGCCGTGATGGCTGGTGACG (SEQ IDNO:537) and 36443r3: GGCAGACTCCTTCCTATGGG (SEQ ID NO:538)] designed fromthe partial human ECE-2 cDNA sequences (DNA49830 and DNA49831). PCRproducts were cloned into the vector pCR2.1-TOPO (Invitrogen Corp.,Carlsbad, Calif., Cat. No. K4500-01) and sequenced with DYE-terminatorchemistry as described above.

Example 98 Northern Blot and in situ RNA Hybridization Analysis forPRO403

[2373] Expression of PRO403 mRNA in human tissues was examined byNorthern blot analysis. Human polyA+ RNA blots derived from human fetaland adult tissues (Clontech, Palo Alto, Calif.; Cat. Nos. 7760-1, 7756-1and 7755-1) were hybridized to a [32P-α]dATP-labeiled cDNA fragmentsfrom probe based on the full length PRO403 cDNA. Blots were incubatedwith the probes in hybridization buffer (5× SSPE; 2× Denhardt'ssolution; 100 mg/mL denatured sheared salmon sperm DNA; 50% formamide;2% SDS) for 18 hours at 42° C., washed to high stringency (0.1× SSC,0.1% SDS, 50° C.) and autoradiographed. The blots were developed afterovernight exposure by phosphorimager analysis (Fuji).

[2374] PRO403 mRNA transcripts were detected. Analysis of the expressionpattern showed the strongest signal of the expected 3.3 kb transcript inadult brain (highest in the cerebellum, putamen, medulla, and temporallobe, and lower in the cerebral cortex, occipital lobe and frontallobe), spinal cord, lung and pancreas and higher levels of a 4.5 kbtranscript in fetal brain and kidney.

Example 99 Use of PRO Polypeptide-Encoding Nucleic Acid as HybridizationProbes

[2375] The following method describes use of a nucleotide sequenceencoding a PRO polypeptide as a hybridization probe.

[2376] DNA comprising the coding sequence of of a PRO polypeptide ofinterest as disclosed herein may be employed as a probe or used as abasis from which to prepare probes to screen for homologous DNAs (suchas those encoding naturally-occurring variants of the PRO polypeptide)in human tissue cDNA libraries or human tissue genomic libraries.

[2377] Hybridization and washing of filters containing either libraryDNAs is performed under the following high stringency conditions.Hybridization of radiolabeled PRO polypeptide-encoding nucleicacid-derived probe to the filters is performed in a solution of 50%formamide, 5× SSC, 0.1% SDS, 0.1% sodium pyrophosphate, 50 mM sodiumphosphate, pH 6.8, 2× Denhardt's solution, and 10% dextran sulfate at42° C. for 20 hours. Washing of the filters is performed in an aqueoussolution of 0.1% SSC and 0.1% SDS at 42° C.

[2378] DNAs having a desired sequence identity with the DNA encodingfull-length native sequence PRO polypeptide can then be identified usingstandard techniques known in the art.

Example 100 Expression of PRO Polypeptides in E. coli

[2379] This example illustrates preparation of an unglycosylated form ofa desired PRO polypeptide by recombinant expression in E. coil.

[2380] The DNA sequence encoding the desired PRO polypeptide isinitially amplified using selected PCR primers. The primers shouldcontain restriction enzyme sites which correspond to the restrictionenzyme sites on the selected expression vector. A variety of expressionvectors may be employed. An example of a suitable vector is pBR322(derived from E. coli; see Bolivar et al., Gene, 2:95 (1977)) whichcontains genes for ampicillin and tetracycline resistance. The vector isdigested with restriction enzyme and dephosphorylated. The PCR amplifiedsequences are then ligated into the vector. The vector will preferablyinclude sequences which encode for an antibiotic resistance gene, a trppromoter, a polyhis leader (including the first six STII codons, polyhissequence, and enterokinase cleavage site), the specific PRO polypeptidecoding region, lambda transcriptional terminator, and an argU gene.

[2381] The ligation mixture is then used to transform a selected E. colistrain using the methods described in Sambrook et al., supra.Transformants are identified by their ability to grow on LB plates andantibiotic resistant colonies are then selected. Plasmid DNA can beisolated and confirmed by restriction analysis and DNA sequencing.

[2382] Selected clones can be grown overnight in liquid culture mediumsuch as LB broth supplemented with antibiotics. The overnight culturemay subsequently be used to inoculate a larger scale culture. The cellsare then grown to a desired optical density, during which the expressionpromoter is turned on.

[2383] After culturing the cells for several more hours, the cells canbe harvested by centriffigation. The cell pellet obtained by thecentrifugation can be solubilized using various agents known in the art,and the solubilized PRO polypeptide can then be purified using a metalchelating column under conditions that allow tight binding of theprotein.

[2384] PRO181, PRO195, PRO200, PRO237, PRO273, PRO540, PRO322, PRO1017,PRO938, PRO162, PRO114, PRO827 and PRO1008 were expressed in E. coli ina poly-His tagged form, using the following procedure. The DNA encodingthe PRO polypeptide was initially amplified using selected PCR primers.The primers contained restriction enzyme sites which correspond to therestriction enzyme sites on the selected expression vector, and otheruseful sequences providing for efficient and reliable translationinitiation, rapid purification on a metal chelation column, andproteolytic removal with enterokinase. The PCR-amplified, poly-Histagged sequences were then ligated into an expression vector, which wasused to transform an E. coli host based on strain 52 (W3110 fuhA (tonA)lon galE rpoHts(htpRts) clpP(lacIq). Transformants were first grown inLB containg 50 mg/ml carbenicillin at 30° C. with shaking until anO.D.600 of 3-5 was reached. Cultures were then diluted 50-100 fold intoCRAP media (prepared by mixing 3.57 g (NH₄)₂SO₄, 0.71 g sodiumcitrate.2H₂O, 1.07 g KCl, 5.36 g Difco yeast extract, 5.36 g Sheffieldhycase SF in 500 mL water, as well as 110 mM MPOS, pH 7.3, 0.55% (w/v)glucose and 7 mM MgSO₄) and grown for approximately 20-30 hours at 30°C. with shaking. Samples were removed to verify expression by SDS-PAGEanalysis, and the bulk culture is centrifuged to pellet the cells. Cellpellets were frozen until purification and refolding.

[2385]E. coli paste from 0.5 to 1 L fermentations (6-10 g pellets) wasresuspended in 10 volumes (w/v) in 7 M guanidine, 20 mM Tris, pH 8buffer. Solid sodium sulfite and sodium tetrathionate is added to makefinal concentrations of 0.1M and 0.02 M, respectively, and the solutionwas stirred overnight at 4° C. This step results in a denatured proteinwith all cysteine residues blocked by sulfitolization. The solution wascentrifuged at 40,000 rpm in a Beckman Ultracentifuge for 30 min. Thesupernatant was diluted with 3-5 volumes of metal chelate column buffer(6 M guanidine, 20 mM Tris, pH 7.4) and filtered through 0.22 micronfilters to clarify. Depending the clarified extract was loaded onto a Sml Qiagen Ni-NTA metal chelate column equilibrated in the metal chelatecolumn buffer. The column was washed with additional buffer containing50 mM imidazole (Calbiochem, Utrol grade), pH 7.4. The protein waseluted with buffer containing 250 mM imidazole. Fractions containing thedesired protein were pooled and stored at 4° C. Protein concentrationwas estimated by its absorbance at 280 nm using the calculatedextinction coefficient based on its amino acid sequence.

[2386] The proteins were refolded by diluting sample slowly into freshlyprepared refolding buffer consisting of: 20 mM Tris, pH 8.6, 0.3 M NaCl,2.5 M urea, 5 mM cysteine, 20 mM glycine and 1 mM EDTA. Refoldingvolumes were chosen so that the final protein concentration was between50 to 100 micrograms/ml. The refolding solution was stirred gently at 4°C. for 12-36 hours. The refolding reaction was quenched by the additionof TFA to a final concentration of 0.4% (pH of approximately 3). Beforefurther purification of the protein, the solution was filtered through a0.22 micron filter and acetonitrile was added to 2-10% finalconcentration. The refolded protein was chromatographed on a Poros RI/Hreversed phase column using a mobile buffer of 0.1% TFA with elutionwith a gradient of acetonitrile from 10 to 80%. Aliquots of fractionswith A280 absorbance were analyzed on SDS polyacrylamide gels andfractions containing homogeneous refolded protein were pooled.Generally, the properly refolded species of most proteins are eluted atthe lowest concentrations of acetonitrile since those species are themost compact with their hydrophobic interiors shielded from interactionwith the reversed phase resin. Aggregated species are usually eluted athigher acetonitrile concentrations. In addition to resolving misfoldedforms of proteins from the desired form, the reversed phase step alsoremoves endotoxin from the samples.

[2387] Fractions containing the desired folded PRO proteins were pooledand the acetonitrile removed using a gentle stream of nitrogen directedat the solution. Proteins were formulated into 20 mM Hepes, pH 6.8 with0.14 M sodium chloride and 4% mannitol by dialysis or by gel filtrationusing G25 Superfine (Pharmacia) resins equilibrated in the formulationbuffer and sterile filtered.

[2388] Many of the PRO polypeptides described herein were successfullyexpressed as described above.

Example 101 Expression of PRO Polypeptides in Mammalian Cells

[2389] This example illustrates preparation of a glycosylated form of adesired PRO polypeptide by recombinant expression in mammalian cells.

[2390] The vector, pRK5 (see EP 307,247, published Mar. 15, 1989), isemployed as the expression vector. Optionally, the PROpolypeptide-encoding DNA is ligated into pRK5 with selected restrictionenzymes to allow insertion of the PRO polypeptide DNA using ligationmethods such as described in Sambrook et al., supra. The resultingvector is called pRK5-PRO polypeptide.

[2391] In one embodiment, the selected host cells may be 293 cells.Human 293 cells (ATCC CCL 1573) are grown to confluence in tissueculture plates in medium such as DMEM supplemented with fetal calf serumand optionally, nutrient components and/or antibiotics. About 10 μgpRK5-PRO polypeptide DNA is mixed with about 1 μg DNA encoding the VARNA gene [Thimmappaya et al., Cell, 31:543 (1982)] and dissolved in 500μl of 1 mM Tris-HCl, 0.1 mM EDTA, 0.227 M CaCl₂. To this mixture isadded, dropwise, 500 μl of 50 mM HEPES (pH 7.35), 280 mM NaCl, 1.5 mMNaPO₄, and a precipitate is allowed to form for 10 minutes at 25° C. Theprecipitate is suspended and added to the 293 cells and allowed tosettle for about four hours at 37° C. The culture medium is aspiratedoff and 2 ml of 20% glycerol in PBS is added for 30 seconds. The 293cells are then washed with serum free medium, fresh medium is added andthe cells are incubated for about 5 days.

[2392] Approximately 24 hours after the transfections, the culturemedium is removed and replaced with culture medium (alone) or culturemedium containing 200 μCi/ml ³⁵S-cysteine and 200 μCi/ml ³⁵S-methionine.After a 12 hour incubation, the conditioned medium is collected,concentrated on a spin filter, and loaded onto a 15% SDS gel. Theprocessed gel may be dried and exposed to film for a selected period oftime to reveal the presence of PRO polypeptide. The cultures containingtransfected cells may undergo further incubation (in serum free medium)and the medium is tested in selected bioassays.

[2393] In an alternative technique, PRO polypeptide may be introducedinto 293 cells transiently using the dextran sulfate method described bySomparyrac et al., Proc. Natl. Acad. Sci., 12:7575 (1981). 293 cells aregrown to maximal density in a spinner flask and 700 μg pRK5-PROpolypeptide DNA is added. The cells are first concentrated from thespinner flask by centrifugation and washed with PBS. The DNA-dextranprecipitate

[2394] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO213-1, PRO1330and/or PRO1449 gene using the probe oligonucleotide and one of the PCRprimers. RNA for construction of the cDNA libraries was isolated fromhuman fetal lung tissue.

[2395] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence encoding PRO213-1, PRO1330 and/or PRO1449(DNA30943-1-1163-1 (SEQ ID NO:505), DNA64907-1163-1 (SEQ ID NO:507) andDNA64908-1163-1 (SEQ ID NO:509), respectively].

[2396] The entire nucleotide sequences corresponding toDNA30943-1-1163-1 (SEQ ID NO:505), DNA64907-1163-1 (SEQ ID NO:507) andDNA64908-1163-1 (SEQ ID NO:509), respectively. DNA30943-1163,DNA64907-1163-1 and DNA64908-1163-1 contain a single open reading framewith an apparent translational initiation site at nucleotide positions336-338, 488-490 and 326-328, respectively, and ending at the stop codonat nucleotide positions 1221-1223, 1307-1309 and 1145-1147, respectively(FIGS. 212, 214 and 216). The predicted polypeptide precursor is 295,273 and 273 amino acids long, respectively (FIGS. 213, 215 and 217).DNA30943-1-1163-1, DNA64907-1163-1 and DNA64908-1163-1 have beendeposited with ATCC and are assigned ATCC deposit no. 209791, 203242 and203243, respectively.

[2397] Analysis of the amino acid sequence of the full-length PRO213-1polypeptide suggests that a portion of it possess significant homologyto the human growth arrest-specific gene 6 protein. More specifically,an analysis of the Dayhoff database (version 35.45 SwissProt 35)evidenced significant homology between the PRO213 amino acid sequenceand the following Dayhoff sequences, HSMHC3W5A_(—)6 and B48089.

[2398] Additional analysis of the amino acid sequence of the full-lengthPRO1330 and PRO1449 polypeptide indicates significant identity withnotch4. More specifically, an analysis of the Dayhoff database (version35.130 SwissProt 35) evidenced significant identity between PRO1330 andthe following Dayhoff sequences, D86566_(—)1 and NEL_HUMAN.

Example 95 Isolation of cDNA Clones Encoding Human PRO298

[2399] A cDNA isolated in the amylase screen described in Example 2above is herein designated DNA26832 (FIG. 220; SEQ ID NO:516). Thesequence of DNA26832 was then used to search expressed sequence tag(EST) databases. The EST databases included public EST databases (e.g.,GenBank) and a proprietary EST database (LIFESEQ™, IncytePharmaceuticals, Palo Alto, Calif.). The search was performed using thecomputer program BLAST or BLAST2 (Altshul et al., Methods in Enzymology266: 469-480 [1996]). Those comparisons resulting in a BLAST score of 70(or in some cases 90) or greater that did not encode proteins wereclustered and assembled into consensus DNA sequences with the program“phrap” (Phil Green, University of Washington, Seattle, Wash.;http:/Ibozeman.mbt.washington.edu/phrap.docs/phrap.html).

[2400] A consensus DNA sequence was assembled relative to other ESTsequences using phrap. A consensus sequence was determined, which wasthen extended using repeated cycles of BLAST and phrap to extend theconsensus sequence as far as possible using the sources of EST sequencesdiscussed above. The extended assembly sequence was designatedDNA35861.Based on the DNA35861 consensus sequence, oligonucleotides weresynthesized: 1) to identity by PCR a cDNA library that contained thesequence of interest, and 2) for use as probes to isolate a clone of thefull-length coding sequence of PRO298. Forward and reverse primersgenerally range from 20 to 30 nucleotides and are often designed to givea PCR product of about 100-1000 bp in length. The probe sequence istypically 40-55 bp in length. In some cases, additional oligonucleotidesare synthesized when the consensus sequence is greater than about 1-1.5kbp. In order to screen several libraries for a full-length clone, DNAfrom the libraries was screened by PCR amplification, as per Ausubel etal., Current Protocols in Molecular Biology, with the PCR primer pair. Apositive library was used to isolate clones encoding the gene ofinterest using the probe oligonucleotide and one of the primer pairs.

[2401] PCR primers (forward and reverse) and a hybridization probe weresynthesized: forward PCR primer 1 CAACGTGATTTCAAAGCTGGGCTC (SEQ IDNO:517) forward PCR primer 2 GCCTCGTATCAAGAATTTCC (SEQ ID NO:518)forward PCR primer 3 AGTGGAAGTCGACCTCCC (SEQ ID NO:519) reverse PCRprimer 1 CTCACCTGAAATCTCTCATAGCCC (SEQ ID NO:520) hybridization probe 1CGCAAAACCCATTTTGGGAGCAGGAATTCCAATCATGTCTGTGATGGTGG (SEQ ID NO:521)

[2402] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO298 gene usingthe probe oligonucleotide and one of the PCR primers.

[2403] RNA for construction of the cDNA libraries was isolated fromhuman fetal lung tissue (LIB25). The cDNA libraries used to isolated thecDNA clones were constructed by standard methods using commerciallyavailable reagents such as those from Invitrogen, San Diego, Calif. ThecDNA was primed with oligo dT containing a Nod site, linked with bluntto Sail hemikinased adaptors, cleaved with NotI, sized appropriately bygel electrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the Sfil site; see, Holmes et al., Science,253:1278-1280 (1991)) in the unique XhoI and NotI sites.

[2404] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO298 (herein designated UNQ261(DNA39975-12101) (SEQ ID NO:514), and the derived protein sequence forPRO298 (SEQ ID NO:515).

[2405] The entire nucleotide sequence of UNQ261 (DNA39975-1210) is shownin FIG. 218 (SEQ ID NO:514). Clone DNA39975-1210 contains a single openreading frame with an apparent translational initiation site atnucleotide positions 375-377. The predicted polypeptide precursor is 364amino acids long. The protein contains four putative transmembranedomains between amino acid positions 36-55 (type II TM), 65-84, 188-208,and 229-245, respectively. A putative N-linked glycosylation site startsat amino acid position 253. In addition, the following features havebeen identified in the protein sequence: cAMP- and cGMP-dependentprotein kinase phosphorylation site, starting at position 8;N-myristoylation sites starting a position 173 and 262, respectively;and a ZP domain between amino acid positions 45-60. Clone DNA39975-1210has been deposited with ATCC (Apr. 21, 1998) and is assigned ATCCdeposit no.209783.

Example 96 Isolation of cDNA Clones Encoding Human PRO337

[2406] A cDNA sequence identified in the amylase screen described inExample 2 above is herein designated DNA42301 (FIG. 223, SEQ ID NO:524).The DNA42301 sequence was then compared to other EST sequences usingphrap as described in Example 1 above and a consensus sequencedesignated herein as DNA28761 was identified. Based on this consensussequence, oligonucleotides were synthesized: 1) to identify -by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence. In orderto screen several libraries for a source of a full-length clone, DNAfrom the libraries was screened by PCR amplification with the PCR primerpair identified above. A positive library was then used to isolateclones encoding the PRO337 gene using the probe oligonucleotide and oneof the PCR primers. RNA for construction of the cDNA libraries wasisolated from human fetal brain.

[2407] A cDNA clone was sequenced in its entirety. The full lengthnucleotide sequence of DNA43316-1237 is shown in FIG. 221 (SEQ IDNO:522). Clone DNA43316-1237 contains a single open reading frame withan apparent translational initiation site at nucleotide positions134-136 (FIG. 221; SEQ ID NO:522). The predicted polypeptide precursoris 344 amino acids long. Clone DNA43316-1237 has been deposited withATCC and is assigned ATCC deposit no. 209487.

[2408] Based on a BLAST-2 and FastA sequence alignment analysis of thefull-length sequence, PRO337 shows amino acid sequence identity to ratneurotimin (97%).

Example 97 Isolation of cDNA Clones Encoding Human PRO403

[2409] Introduction:

[2410] Human thrombopoietin (THPO) is a glycosylated hormone of 352amino acids consisting of two domains. The N-terminal domain, sharing50% similarity to erythropoietin, is responsible for the biologicalactivity. The C-terminal region is required for secretion. The gene forthrombopoietin (ThPO) maps to human chromosome 3q27-q8 where the sixexons of this gene span 7 kilobase base pairs of genomic DNA (Chang etal., Genomics 26: 636-7 (1995); Foster et al., Proc. Natl. Acad. Sci.USA 91: 13023-7 (1994); Gurney et al., Blood 85: 981-988 (1995). Inorder to determine whether there were any genes encoding THPO homologueslocated in close proximity to THPO, genomic DNA fragments from thisregion were identified and sequenced. Three P1 clones and one PAC clones(Genome Systems Inc., St. Louis, Mo.; cat. Nos. P1-2535 and PAC-6539)encompassing the THPO locus were isolated and a 140 kb region wassequenced using the ordered shotgun strategy (Chenet al., Genomics 17:651-656 (1993)), coupled with a PCR-based gap filling approach. Analysisreveals that the region is gene-rich with four additional genes locatedvery close to THPO: tumor necrosis factor- receptor type 1 associatedprotein 2 (TRAP2) and elongation initiation factor gamma (elF4( ),chloride channel 2 (CLCN2) and RNA polymerase II subunit hRPB17. Whileno THPO homolog was found in the region, four novel genes have beenpredicted by computer-assisted gene detection (GRAIL)(Xu et al., Gen.Engin. 16: 241-253 (1994), the presence of CpG islands (Cross, S. andBird, A., Curr. Opin. Genet. & Devel. 5: 109-314 (1995), and homology toknown genes (as detected by WU-BLAST2.0)(Altschul and Gish, MethodsEnzymol. 266: 460-480 (1996) (http://blast.wustl.edu/blast/README.html).

[2411] Procedures:

[2412] P1 and PAC Clones:

[2413] The initial human P1 clone was isolated from a genomic P1 library(Genome Systems Inc., St. Louis, MO; cat. no.: P1-2535) screened withPCR primers designed from the THPO genomic sequence (A. L. Gurney, etal., Blood 85: 981-88 (1995). PCR primers were designed from the endsequences derived from this P1 clone were then used to screen P1 and PAClibraries (Genome Systems, Cat. Nos.: P1-2535 & PAC-6539) to identifyoverlapping clones (PAC1, p1.t, and P1.u). The 3′-end sequence fromPAC.z was used to define the primers used for the screening of a humanBAC library (Genome Systems Inc., St. Louis, MO; Cat. No.: BDTW-4533A).

[2414] Ordered Shotgtun Strategy:

[2415] The Ordered Shotgun Strategy (OSS) (Chen et al., Genomics 17:651-656 (1993)) involves the mapping and sequencing of large genomic DNAclones with a hierarchical approach. The P1 or PAC clone was sonicatedand the fragments subcloned into lambda vector (ABluestar) (Novagen,Inc., Madison, Wis.; cat. no. 69242-3). The lambda subclone inserts wereisolated by long-range PCR (Barnes, W. Proc. Natl. Acad. Sci. USA 91:2216-2220 (1994) and the ends sequenced. The lambda-end sequences wereoverlapped to create a partial map of the original clone. Those lambdaclones with overlapping end-sequences were identified, the insetssubcloned into a plasmid vector (pUC18 or pUC19, Hoefer PharmaciaBiotech, Inc., San Francisco, Calif., Cat. Nos. 27-4949-01 and274951-01) and the ends of the plasmid subclones were sequenced andassembled to generate a contiguous sequence. This directed sequencingstrategy minimizes the redundancy required while allowing one to scanfor and concentrate on interesting regions.

[2416] In order to define better the THPO locus and to search for othergenes related to the hematopoietin family, five genomic clones wereisolated from this region by PCR screening of human PI and PAC libraries(Genome System, Inc., Cat. Nos.: P1-2535 and PAC-6539). _os The sizes ofthe genomic fragments are as follows: P1.t is 40 kb; P1.g is 70 kb; P1.uis 70 kb; PAC.z is 200 kb; and BAC.1 is 80 kb. Approximately 75% (140kb) of the 190 kb genomic DNA region was sequenced by the OrderedShotgun Strategy (OSS) (Chen et al., Genomics 17: 651-56 (1993), andassembled into contigs using AutoAssemblerTM (Applied Biosystems, PerkinElmer, Foster City, Calif., cat. no. 903227). The preliminary order ofthese contigs was determined by manual analysis. There were 47 contigsthe 140 kb region. A PCR-based approach to ordering the contigs andfilling in the gaps was employed. The following summarizes the numberand sizes of the gaps. The 50 kb of sequence unique to BAC.1 wassequenced by a total shotgun approach with a ten-fold redundancy. Sizeof gap number <50 bp 13  50-150 bp  7  150-300 bp  7  300-1000 bp 101000-5000 bp  7 >5000 bp  2 ((15,000 bp)

[2417] DNA Sequencing:

[2418] ABI DYE-primerTM chemistry (PE Applied Biosystems, Foster City,Calif.; Cat. No.: 402112) was used to end-sequence the lambda andplasmid subclones. ABI DYE-terminaterTM chemistry (PE AppliedBiosystems, Foster City, Calif., Cat. No: 403044) was used to sequencethe PCR products with their respective PCR primers. The sequences werecollected with an ABI377 instrument. For PCR products larger than 1 kb,walking primers were used. The sequences of contigs generated by the OSSstrategy in AutoAssemblerTM (PE Applied Biosystems, Foster City, Calif.;Cat. No: 903227) and the gap-filling sequencing trace files wereimported into SequencherTM (Gene Codes Corp., Ann Arbor, Mich.) foroverlapping and editing. The sequences generated by the total shotgunstrategy were assembled using Phred and Phrap and edited using Consed(http://chimera.biotech.washington.eduluwgc/projects.htm) and GFP(Genome Reconstruction Manager for Phrap), version 1.2(http://stork.cellb.bcm.tmc.edu/gfp/).

[2419] PCR-Based Gap Filling Strategy:

[2420] Primers were designed based on the 5′- and 3′-end sequenced ofeach contig, avoiding repetitive and low quality sequence regions. Allprimers were designed to be 19-24-mers with 50-70% G/C content. Oligoswere synthesized and gel-purified by standard methods.

[2421] Since the orientation and order of the contigs were unknown,permutations of the primers were used in the amplification reactions.Two PCR kits were used: first, XL PCR kit (Perkin Elmer, Norwalk, Conn.;Cat. No.: N8080205), with extension times of approximately 10 minutes;and second, the Taq polymerase PCR kit (Qiagen Inc., Valencia, Calif.;Cat. No.: 201223) was used under high stringency conditions if smearedor multiple products were observed with the XL PCR kit. The main PCRproduct from each successful reaction was extracted from a 0.9% lowmelting agarose gel and purified with the Geneclean DNA Purification kitprior to sequencing.

[2422] Analysis:

[2423] The identification and characterization of coding regions wascarried out as follows: First, repetitive sequences were masked usingRepeatMasker (A. F. A. Smit & P. Green, http://flp.genome.washington.eduRM/RM_details.html) which screens DNA sequences in FastA format againsta library of repetitive elements and returns a masked query sequence.Repeats not masked were identified by comparing the sequence to theGenBank database using WUBLAST2.0 [Altschul, S & Gish, W., MethodsEnzymol. 266: 460-480 (1996); http://blast.wustl.edu/blast/README.html]and were masked manually.

[2424] Next, known genes were revealed by comparing the genomic regionsagainst Genentech's protein database using the WUBLAST2.0 algorithm andthen annotated by aligning the genomic and cDNA sequences for each gene,respectively, using a Needleman-Wunch (Needleman and Wunsch, J. Mol.Biol. 48: 443-453 (1970) algorithm to find regions of local identitybetween sequences. The strategy results in detection of all exons of thefive known genes in the region, THPO, TRAP2, eIF4g, CLCN2 and hRPB17(see below). Known genes Map position eukaryotic translation initiationfactor 4 gamma 3q27-qter thrombopoietin 3q26-q27 chloride channel 23q26-qter TNF receptor associated protein 2 not previously mapped RNApolymerase II subunit hRPB17 not previously mapped

[2425] Finally, novel transcription units were predicted using a numberof approaches. CpG islands (S. Cross & Bird, A., Curr. Opin. Genet. Dev.5: 109-314 (1995) islands were used to define promoter regions and wereidentified as clusters of sites cleaved by enzymes recognizing GC-rich,6 or 8-mer palindromic sequences (NotI, NarI, BssHII, XhoI. CpG islandsare usually associated with promoter regions of genes. WUBLAST2.0analysis of short genomic regions (10-20 kb) versus GenBank revealedmatches to ESTs. The individual EST sequences (or where possible, theirsequence chromatogram files) were retrieved and assembled with Sequencerto provide a theoretical cDNA sequence (DNA36443). GRAIL2 (ApoCom Inc.,Knoxville, Tenn., command line version for the DEC alpha) was used topredict a novel exon. The five known genes in the region served asinternal controls for the success of the GRAIL algorithm.

[2426] Isolation:

[2427] A partial endothelin converting enzyme-2 (ECE-2) cDNA clone wasisolated by first splicing in silico the ECE-2 exons predicted in thegenomic sequence to generate a putative sequence (DNA36443). Anoligonucleotide probe: GAAGCAGTGCAGCCAGCAGTAGAGAGGCACCTGCTAAGA) (SEQ IDNO:530) was designed and used to screen a human fetal small intestinelibrary (LIB110) and internal PCR primers (36443fl)(ECE2.f:ACGCAGCTGGAGCTGGTCTTAGCA) (SEQ ID NO:531) and (36443r1) (ECE2.r)(GGTACTGGACCCCTAGGGCCACAA) (SEQ ID NO:532) were used to confirm cloneshybridizing to the probe prior to sequencing. One positive clone wasobtained, however this cDNA (DNA49830) represented a partially splicedtranscript containing appropriately spliced exons 1 through 6, followedby intron 6 sequence. The oligo dT primer annealed to a polyA-stretchwithin an Alu element present in intron 6. An additional ECE-2 cDNAfragment (DNA49831) was obtained by PCR from a human fetal kidneylibrary (LIB227) with primers designed from the presumed cDNA sequence[36443f3: CCTCCCAGCCGAGACCAGTGG (SEQ ID NO:533) and 36443r2:GGTCCTATAAGGGCCAAGACC (SEQ ID NO:534)]. This PCR product extended fromexon 13 into the 3′ untranslated region in exon 18.

[2428] A full length endothelin converting enzyme 2 (ECE-2) cDNA clone(DNA55800-1263) was isolated from an oligo-dT-primed human fetal brainlibrary. RNA from human fetal brain tissue (20 weeks gestation,#283005)(SRC175) was isolated by guanidine thiocyanate and 5 μg used togenerate double stranded cDNA which was cloned into the vector pRK5E.The 3′-primer (pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTTT) (SEQ IDNO:535) and the 5-linker (pCGGACGCGTGGGTCGA) (SEQ ID NO:536) weredesigned to introduce XhoI and NotI restriction sites. The library wasscreened with PCR primers [36443pcrf1: CGGCCGTGATGGCTGGTGACG (SEQ IDNO:537) and 36443r3: GGCAGACTCCTTCCTATGGG (SEQ ID NO:538)] designed fromthe partial human ECE-2 cDNA sequences (DNA49830 and DNA49831). PCRproducts were cloned into the vector pCR2.1-TOPO (Invitrogen Corp.,Carlsbad, Calif., Cat. No. K4500-01) and sequenced with DYE-terminatorchemistry as described above.

Example 98 Northern Blot and in situ RNA Hybridization Analysis forPRO403

[2429] Expression of PRO403 mRNA in human tissues was examined byNorthern blot analysis. Human polyA+ RNA blots derived from human fetaland adult tissues (Clontech, Palo Alto, Calif.; Cat. Nos. 7760-1, 7756-1and 7755-1) were hybridized to a [32P-α]dATP-labeiled cDNA fragmentsfrom probe based on the full length PRO403 cDNA. Blots were incubatedwith the probes in hybridization buffer (5× SSPE; 2× Denhardt'ssolution; 100 mg/mL denatured sheared salmon sperm DNA; 50% formamide;2% SDS) for 18 hours at 42° C., washed to high stringency (0.1× SSC,0.1% SDS, 50° C.) and autoradiographed. The blots were developed afterovernight exposure by phosphorimager analysis (Fuji).

[2430] PRO403 mRNA transcripts were detected. Analysis of the expressionpattern showed the strongest signal of the expected 3.3 kb transcript inadult brain (highest in the cerebellum, putamen, medulla, and temporallobe, and lower in the cerebral cortex, occipital lobe and frontallobe), spinal cord, lung and pancreas and higher levels of a 4.5 kbtranscript in fetal brain and kidney.

Example 99 Use of PRO Polypeptide-Encoding Nucleic Acid as HybridizationProbes

[2431] The following method describes use of a nucleotide sequenceencoding a PRO polypeptide as a hybridization probe.

[2432] DNA comprising the coding sequence of of a PRO polypeptide ofinterest as disclosed herein may be employed as a probe or used as abasis from which to prepare probes to screen for homologous DNAs (suchas those encoding naturally-occurring variants of the PRO polypeptide)in human tissue cDNA libraries or human tissue genomic libraries.

[2433] Hybridization and washing of filters containing either libraryDNAs is performed under the following high stringency conditions.Hybridization of radiolabeled PRO polypeptide-encoding nucleicacid-derived probe to the filters is performed in a solution of 50%formamide, 5× SSC, 0.1% SDS, 0.1% sodium pyrophosphate, 50 mM sodiumphosphate, pH 6.8, 2× Denhardt's solution, and 10% dextran sulfate at42° C. for 20 hours. Washing of the filters is performed in an aqueoussolution of 0.1% SSC and 0.1% SDS at 42° C.

[2434] DNAs having a desired sequence identity with the DNA encodingfull-length native sequence PRO polypeptide can then be identified usingstandard techniques known in the art.

Example 100 Expression of PRO Polypeptides in E. coli

[2435] This example illustrates preparation of an unglycosylated form ofa desired PRO polypeptide by recombinant expression in E. coil.

[2436] The DNA sequence encoding the desired PRO polypeptide isinitially amplified using selected PCR primers. The primers shouldcontain restriction enzyme sites which correspond to the restrictionenzyme sites on the selected expression vector. A variety of expressionvectors may be employed. An example of a suitable vector is pBR322(derived from E. coli; see Bolivar et al., Gene, 2:95 (1977)) whichcontains genes for ampicillin and tetracycline resistance. The vector isdigested with restriction enzyme and dephosphorylated. The PCR amplifiedsequences are then ligated into the vector. The vector will preferablyinclude sequences which encode for an antibiotic resistance gene, a trppromoter, a polyhis leader (including the first six STII codons, polyhissequence, and enterokinase cleavage site), the specific PRO polypeptidecoding region, lambda transcriptional terminator, and an argU gene.

[2437] The ligation mixture is then used to transform a selected E. colistrain using the methods described in Sambrook et al., supra.Transformants are identified by their ability to grow on LB plates andantibiotic resistant colonies are then selected. Plasmid DNA can beisolated and confirmed by restriction analysis and DNA sequencing.

[2438] Selected clones can be grown overnight in liquid culture mediumsuch as LB broth supplemented with antibiotics. The overnight culturemay subsequently be used to inoculate a larger scale culture. The cellsare then grown to a desired optical density, during which the expressionpromoter is turned on.

[2439] After culturing the cells for several more hours, the cells canbe harvested by centriffigation. The cell pellet obtained by thecentrifugation can be solubilized using various agents known in the art,and the solubilized PRO polypeptide can then be purified using a metalchelating column under conditions that allow tight binding of theprotein.

[2440] PRO181, PRO195, PRO200, PRO237, PRO273, PRO540, PRO322, PRO1017,PRO938, PRO162, PRO114, PRO827 and PRO1008 were expressed in E. coli ina poly-His tagged form, using the following procedure. The DNA encodingthe PRO polypeptide was initially amplified using selected PCR primers.The primers contained restriction enzyme sites which correspond to therestriction enzyme sites on the selected expression vector, and otheruseful sequences providing for efficient and reliable translationinitiation, rapid purification on a metal chelation column, andproteolytic removal with enterokinase. The PCR-amplified, poly-Histagged sequences were then ligated into an expression vector, which wasused to transform an E. coli host based on strain 52 (W3110 fuhA (tonA)lon galE rpoHts(htpRts) clpP(lacIq). Transformants were first grown inLB containg 50 mg/ml carbenicillin at 30° C. with shaking until anO.D.600 of 3-5 was reached. Cultures were then diluted 50-100 fold intoCRAP media (prepared by mixing 3.57 g (NH₄)₂SO₄, 0.71 g sodiumcitrate.2H₂O, 1.07 g KCl, 5.36 g Difco yeast extract, 5.36 g Sheffieldhycase SF in 500 mL water, as well as 110 mM MPOS, pH 7.3, 0.55% (w/v)glucose and 7 mM MgSO₄) and grown for approximately 20-30 hours at 30°C. with shaking. Samples were removed to verify expression by SDS-PAGEanalysis, and the bulk culture is centrifuged to pellet the cells. Cellpellets were frozen until purification and refolding.

[2441]E. coli paste from 0.5 to 1 L fermentations (6-10 g pellets) wasresuspended in 10 volumes (w/v) in 7 M guanidine, 20 mM Tris, pH 8buffer. Solid sodium sulfite and sodium tetrathionate is added to makefinal concentrations of 0.1M and 0.02 M, respectively, and the solutionwas stirred overnight at 4° C. This step results in a denatured proteinwith all cysteine residues blocked by sulfitolization. The solution wascentrifuged at 40,000 rpm in a Beckman Ultracentifuge for 30 min. Thesupernatant was diluted with 3-5 volumes of metal chelate column buffer(6 M guanidine, 20 mM Tris, pH 7.4) and filtered through 0.22 micronfilters to clarify. Depending the clarified extract was loaded onto a Sml Qiagen Ni-NTA metal chelate column equilibrated in the metal chelatecolumn buffer. The column was washed with additional buffer containing50 mM imidazole (Calbiochem, Utrol grade), pH 7.4. The protein waseluted with buffer containing 250 mM imidazole. Fractions containing thedesired protein were pooled and stored at 4° C. Protein concentrationwas estimated by its absorbance at 280 nm using the calculatedextinction coefficient based on its amino acid sequence.

[2442] The proteins were refolded by diluting sample slowly into freshlyprepared refolding buffer consisting of: 20 mM Tris, pH 8.6, 0.3 M NaCl,2.5 M urea, 5 mM cysteine, 20 mM glycine and 1 mM EDTA. Refoldingvolumes were chosen so that the final protein concentration was between50 to 100 micrograms/ml. The refolding solution was stirred gently at 4°C. for 12-36 hours. The refolding reaction was quenched by the additionof TFA to a final concentration of 0.4% (pH of approximately 3). Beforefurther purification of the protein, the solution was filtered through a0.22 micron filter and acetonitrile was added to 2-10% finalconcentration. The refolded protein was chromatographed on a Poros RI/Hreversed phase column using a mobile buffer of 0.1% TFA with elutionwith a gradient of acetonitrile from 10 to 80%. Aliquots of fractionswith A280 absorbance were analyzed on SDS polyacrylamide gels andfractions containing homogeneous refolded protein were pooled.Generally, the properly refolded species of most proteins are eluted atthe lowest concentrations of acetonitrile since those species are themost compact with their hydrophobic interiors shielded from interactionwith the reversed phase resin. Aggregated species are usually eluted athigher acetonitrile concentrations. In addition to resolving misfoldedforms of proteins from the desired form, the reversed phase step alsoremoves endotoxin from the samples.

[2443] Fractions containing the desired folded PRO proteins were pooledand the acetonitrile removed using a gentle stream of nitrogen directedat the solution. Proteins were formulated into 20 mM Hepes, pH 6.8 with0.14 M sodium chloride and 4% mannitol by dialysis or by gel filtrationusing G25 Superfine (Pharmacia) resins equilibrated in the formulationbuffer and sterile filtered.

[2444] Many of the PRO polypeptides described herein were successfullyexpressed as described above.

Example 101 Expression of PRO Polypeptides in Mammalian Cells

[2445] This example illustrates preparation of a glycosylated form of adesired PRO polypeptide by recombinant expression in mammalian cells.

[2446] The vector, pRK5 (see EP 307,247, published Mar. 15, 1989), isemployed as the expression vector. Optionally, the PROpolypeptide-encoding DNA is ligated into pRK5 with selected restrictionenzymes to allow insertion of the PRO polypeptide DNA using ligationmethods such as described in Sambrook et al., supra. The resultingvector is called pRK5-PRO polypeptide.

[2447] In one embodiment, the selected host cells may be 293 cells.Human 293 cells (ATCC CCL 1573) are grown to confluence in tissueculture plates in medium such as DMEM supplemented with fetal calf serumand optionally, nutrient components and/or antibiotics. About 10 μgpRK5-PRO polypeptide DNA is mixed with about 1 μg DNA encoding the VARNA gene [Thimmappaya et al., Cell, 31:543 (1982)] and dissolved in 500μl of 1 mM Tris-HCl, 0.1 mM EDTA, 0.227 M CaCl₂. To this mixture isadded, dropwise, 500 μl of 50 mM HEPES (pH 7.35), 280 mM NaCl, 1.5 mMNaPO₄, and a precipitate is allowed to form for 10 minutes at 25° C. Theprecipitate is suspended and added to the 293 cells and allowed tosettle for about four hours at 37° C. The culture medium is aspiratedoff and 2 ml of 20% glycerol in PBS is added for 30 seconds. The 293cells are then washed with serum free medium, fresh medium is added andthe cells are incubated for about 5 days.

[2448] Approximately 24 hours after the transfections, the culturemedium is removed and replaced with culture medium (alone) or culturemedium containing 200 μCi/ml ³⁵S-cysteine and 200 μCi/ml ³⁵S-methionine.After a 12 hour incubation, the conditioned medium is collected,concentrated on a spin filter, and loaded onto a 15% SDS gel. Theprocessed gel may be dried and exposed to film for a selected period oftime to reveal the presence of PRO polypeptide. The cultures containingtransfected cells may undergo further incubation (in serum free medium)and the medium is tested in selected bioassays.

[2449] In an alternative technique, PRO polypeptide may be introducedinto 293 cells transiently using the dextran sulfate method described bySomparyrac et al., Proc. Natl. Acad. Sci., 12:7575 (1981). 293 cells aregrown to maximal density in a spinner flask and 700 μg pRK5-PROpolypeptide DNA is added. The cells are first concentrated from thespinner flask by centrifugation and washed with PBS. The DNA-dextranprecipitate is incubated on the cell pellet for four hours. The cellsare treated with 20% glycerol for 90 seconds, washed with tissue culturemedium, and re-introduced into the spinner flask containing tissueculture medium, 5 μg/ml bovine insulin and 0.1 μg/ml bovine transferrin.After about four days, the conditioned media is centrifuged and filteredto remove cells and debris. The sample containing expressed PROpolypeptide can then be concentrated and purified by any selectedmethod, such as dialysis and/or column chromatography.

[2450] In another embodiment, PRO polypeptides can be expressed in CHOcells. The pRK5-PRO polypeptide can be transfected into CHO cells usingknown reagents such as CaPO₄ or DEAE-dextran. As described above, thecell cultures can be incubated, and the medium replaced with culturemedium (alone) or medium containing a radiolabel such as ³⁵S-methionine.After determining the presence of PRO polypeptide, the culture mediummay be replaced with serum free medium. Preferably, the cultures areincubated for about 6 days, and then the conditioned medium isharvested. The medium containing the expressed PRO polypeptide can thenbe concentrated and purified by any selected method.

[2451] Epitope-tagged PRO polypeptide may also be expressed in host CHOcells. The PRO polypeptide may be subcloned out of the pRK5 vector. Thesubclone insert can undergo PCR to fuse in frame with a selected epitopetag such as a poly-his tag into a Baculovirus expression vector. Thepoly-his tagged PRO polypeptide insert can then be subcloned into a SV40driven vector containing a selection marker such as DHFR for selectionof stable clones. Finally, the CHO cells can be transfected (asdescribed above) with the SV40 driven vector. Labeling may be performed,as described above, to verify expression. The culture medium containingthe expressed poly-His tagged PRO polypeptide can then be concentratedand purified by any selected method, such as by Ni²⁺-chelate affinitychromatography.

[2452] Stable expression in CHO cells was performed using the followingprocedure. The proteins were expressed as an IgG construct(immunoadhesin), in which the coding sequences for the soluble forms(e.g. extracellular domains) of the respective proteins were fused to anIgG1 constant region sequence containing the hinge, CH2 and CH2 domainsand/or is a poly-His tagged form.

[2453] Following PCR amplification, the respective DNAs were subclonedin a CHO expression vector using standard techniques as described inAusubel et al., Current Protocols of Molecular Biology, Unit 3.16, JohnWiley and Sons (1997). CHO expression vectors are constructed to havecompatible restriction sites 5′ and 3′ of the DNA of interest to allowthe convenient shuttling of cDNA's. The vector used expression in CHOcells is as described in Lucas et al., Nucl. Adds Res. 24: 9 (1774-1779(1996), and uses the SV40 early promoter/enhancer to drive expression ofthe cDNA of interest and dihydrofolate reductase (DHFR). DHFR expressionpermits selection for stable maintenance of the plasmid followingtransfection.

[2454] Twelve micrograms of the desired plasmid DNA were introduced intoapproximately 10 million CHO cells using commercially availabletransfection reagents Superfect® (Quiagen), Dosper® or Eugene®(Boehringer Mannheim). The cells were grown and described in Lucas etal., supra. Approximately 3×10⁻⁷ cells are frozen in an ampule forfurther growth and production as described below.

[2455] The ampules containing the plasmid DNA were thawed by placementinto water bath and mixed by vortexing. The contents were pipetted intoa centrifuge tube containing 10 mLs of media and centrifuged at 1000 rpmfor 5 minutes. The supernatant was aspirated and the cells wereresuspended in 10 mL of selective media (0.2 μm filtered PS20 with 5%0.2 lm diafiltered fetal bovine serum). The cells were then aliquotedinto a 100 mL spinner containing 90 mL of selective media. After 1-2days, the cells were transferred into a 250 mL spinner fled with 150 mLselective growth medium and incubated at 37° C. After another 2-3 days,a 250 mL, 500 mL and 2000 mL spinners were seeded with 3×10⁵ cells/mL.The cell media was exchanged with fresh media by centrifugation andresuspension in production medium. Although any suitable CHO media maybe employed, a production medium described in U.S. Pat. No. 5,122,469,issued Jun. 16, 1992 was actually used. 3L production spinner is seededat 1.2×10⁶ cells/mL. On day 0, the cell number pH were determined. Onday 1, the spinner was sampled and sparging with filtered air wascommenced. On day 2, the spinner was sampled, the temperature shifted to33° C., and 30 mL of 500 g/L glucose and 0.6 mL of 10% antifoam (e.g.,35% polydimethylsiloxane emulsion, Dow Corning 365 Medical GradeEmulsion). Throughout the production, pH was adjusted as necessary tokeep at around 7.2. After 10 days, or until viability dropped below 70%,the cell culture was harvested by centrifugtion and filtering through a0.22 μm filter. The filtrate was either stored at 4° C. or immediatelyloaded onto columns for purification.

[2456] For the poly-His tagged constructs, the proteins were purifiedusing a Ni-NTA column (Qiagen). Before purification, imidazole was addedto the conditioned media to a concentration of 5 mM. The conditionedmedia was pumped onto a 6 ml Ni-NTA column equilibrated in 20 mM Hepes,pH 7.4, buffer containing 0.3 M NaCl and 5 mM imidazole at a flow rateof 4-5 ml/min. at 4° C. After loading, the column was washed withadditional equilibration buffer and the protein eluted withequilibration buffer containing 0.25 M imidazole. The highly purifiedprotein was subsequently desalted into a storage buffer containing 10 mMHepes, 0.14 M NaCl and 4% mannitol, pH 6.8, with a 25 ml G25 Superfine(Pharmacia) column and stored at −80° C.

[2457] Immunoadhesin (Fc containing) constructs of were purified fromthe conditioned media as follows. The conditioned medium was pumped ontoa 5 ml Protein A column (Pharmacia) which had been equilibrated in 20 mMNa phosphate buffer, pH 6.8. After loading, the column was washedextensively with equilibration buffer before elution with 100 mM citricacid, pH 3.5. The eluted protein was immediately neutralized bycollecting 1 ml fractions into tubes containing 275 μL of 1 M Trisbuffer, pH 9. The highly purified protein was subsequently desalted intostorage buffer as described above for the poly-His tagged proteins. Thehomogeneity was assessed by SDS polyacrylamide gels and by N-terminalamino acid sequencing by Edman degradation.

[2458] Many of the PRO polypeptides described herein were successfuillyexpressed as described above.

Example 102 Expression of PRO Polypeptides in Yeast

[2459] The following method describes recombinant expression of adesired PRO polypeptide in yeast.

[2460] First, yeast expression vectors are constructed for intracellularproduction or secretion of PRO polypeptides from the ADH2/GAPDHpromoter. DNA encoding a desired PRO polypeptide, a selected signalpeptide and the promoter is inserted into suitable restriction enzymesites in the selected plasmid to direct intracellular expression of thePRO polypeptide. For secretion, DNA encoding the PRO polypeptide can becloned into the selected plasmid, together with DNA encoding theADH2/GAPDH promoter, the yeast alpha-factor secretory signal/leadersequence, and linker sequences (if needed) for expression of the PROpolypeptide.

[2461] Yeast cells, such as yeast strain AB110, can then be transformedwith the expression plasmids described above and cultured in selectedfermentation media. The transformed yeast supernatants can be analyzedby precipitation with 10% trichloroacetic acid and separation bySDS-PAGE, followed by staining of the gels with Coomassie Blue stain.

[2462] Recombinant PRO polypeptide can subsequently be isolated andpurified by removing the yeast cells from the fermentation medium bycentrigation and then concentrating the medium using selected cartridgefilters. The concentrate containing the PRO polypeptide may further bepurified using selected column chromatography resins.

[2463] Many of the PRO polypeptides described herein were successfuillyexpressed as described above.

Example 103 Expression of PRO Polypeptides in Baculovirus-InfectedInsect Cells

[2464] The following method describes recombinant expression of PROpolypeptides in Baculovirus-infected insect cells.

[2465] The desired PRO polypeptide is fused upstream of an epitope tagcontained with a baculovirus expression vector. Such epitope tagsinclude poly-his tags and immunoglobulin tags (like Fc regions of IgG).A variety of plasmids may be employed, including plasmids derived fromcommercially available plasmids such as pVL1393 (Novagen). Briefly, thePRO polypeptide or the desired portion of the PRO polypeptide (such asthe sequence encoding the extracellular domain of a transmembraneprotein) is amplified by PCR with primers complementary to the 5′ and 3′regions. The 5′ primer may incorporate flanking (selected) restrictionenzyme sites. The product is then digested with those selectedrestriction enzymes and subcloned into the expression vector.

[2466] Recombinant baculovirus is generated by co-transfecting the aboveplasmid and BaculoGold™ virus DNA (Pharmingen) into Spodopterafrugiperda (“Sf9”) cells (ATCC CRL 1711) using lipofectin (commerciallyavailable from GIBCO-BRL). After 4-5 days of incubation at 28° C., thereleased viruses are harvested and used for further amplifications.Viral infection and protein expression is performed as described byO'Reilley et al., Baculovirus expression vectors: A laboratory Manual,Oxford: Oxford University Press (1994).

[2467] Expressed poly-his tagged PRO polypeptide can then be purified,for example, by Ni²⁺-chelate affinity chromatography as follows.Extracts are prepared from recombinant virus-infected Sf9 cells asdescribed by Rupert et al., Nature, 362:175-179 (1993). Briefly, Sf9cells are washed, resuspended in sonication buffer (25 mL Hepes, pH 7.9;12.5 mM MgCl₂; 0.1 mM EDTA; 10% Glycerol; 0.1% NP40; 0.4 M KCl), andsonicated twice for 20 seconds on ice. The sonicates are cleared bycentrifugation, and the supernatant is diluted 50-fold in loading buffer(50 mM phosphate, 300 mM NaCl, 10% Glycerol, pH 7.8) and filteredthrough a 0.45 μm filter. A Ni²⁺-NTA agarose column (commerciallyavailable from Qiagen) is prepared with a bed volume of S nL, washedwith 25 mL of water and equilibrated with 25 mL of loading buffer. Thefiltered cell extract is loaded onto the column at 0.5 mL per minute.The column is washed to baseline A₂₈₀ with loading buffer, at whichpoint fraction collection is started. Next, the column is washed with asecondary wash buffer (50 mM phosphate; 300 mM NaCl, 10% Glycerol, pH6.0), which elutes nonspecifically bound protein. After reaching A2mbaseline again, the column is developed with a 0 to 500 mM Imidazolegradient in the secondary wash buffer. One mL fractions are collectedand analyzed by SDS-PAGE and silver staining or western blot withNi²⁺-NTA-conjugated to alkaline phosphatase (Qiagen). Fractionscontaining the eluted His₁₀-tagged PRO polypeptide are pooled anddialyzed against loading buffer.

[2468] Alternatively, purification of the IgG tagged (or Fc tagged) PROpolypeptide can be performed using known chromatography techniques,including for instance, Protein A or protein G column chromatography.PRO195, PRO526, PRO540, PRO846, PRO362, PRO363, PRO700, PRO707, PRO322,PRO719, PRO1083, PRO868, PRO866, PRO768, PRO788, PRO938, PRO827 andPRO1031 were successfully expressed in baculovirus infected Sf9 insectcells. While the expression was actually performed in a 0.5-2 L scale,it can be readily scaled up for larger (e.g. 8 L) preparations. Theproteins were expressed as an IgG construct (immunoadhesin), in whichthe protein extracellular region was fused to an IgG1 constant regionsequence containing the hinge, CH2 and CH3 domains and/or in poly-Histagged forms.

[2469] For expression in baculovirus infected Sf9 cells, following PCRamplification, the respective coding sequences were subcloned into abaculovirus expression vector (pb.PH.IgG for IgG fusions and pb.PH.His.cfor poly-His tagged proteins), and the vector and Baculogold®baculovirus DNA (Pharmingen) were co-transfected into 105 Spodopterafrugiperda (“Sf9”) cells (ATCC CRL 1711), using Lipofectin (Gibco BRL).pb.PH.IgG and pb.PH.His are modifications of the commercially availablebaculovirus expression vector pVL1393 (Pharmingen), with modifiedpolylinker regions to include the His or Fc tag sequences. The cellswere grown in Hink's TNM-FH medium supplemented with 10% FBS (Hyclone).Cells were incubated for 5 days at 28° C. The supernatant was harvestedand subsequently used for the first viral amplification by infecting Sf9cells in Hink's TNM-FH medium supplemented with 10% FBS at anapproximate multiplicity of infection (MOI) of 10. Cells were incubatedfor 3 days at 28° C. The supernatant was harvested and the expression ofthe constructs in the baculovirus expression vector was determined bybatch binding of 1 ml of supernatant to 25 mL of Ni-NTA beads (QIAGEN)for histidine tagged proteins or Protein-A Sepharose CL-4B beads(Pharmacia) for IgG tagged proteins followed by SDS-PAGE analysiscomparing to a known concentration of protein standard by Coomassie bluestaining.

[2470] The first viral amplification supernatant was used to infect aspinner culture (500 ml) of Sf9 cells grown in ESF-921 medium(Expression Systems LLC) at an approximate MOI of 0.1. Cells wereincubated for 3 days at 28° C. The supernatant was harvested andfiltered. Batch binding and SDS-PAGE analysis was repeated, asnecessary, until expression of the spinner culture was confirmed.

[2471] The conditioned medium from the transfected cells (0.5 to 3 L)was harvested by centrifugation to remove the cells and filtered through0.22 micron filters. For the poly-His tagged constructs, the proteinconstruct were purified using a Ni-NTA column (Qiagen). Beforepurification, imidazole was added to the conditioned media to aconcentration of 5 mM. The conditioned media were pumped onto a 6 mlNi-NTA column equilibrated in 20 mM Hepes, pH 7.4, buffer containing 0.3M NaCl and 5 mM imidazole at a flow rate of 4-5 ml/min. at 4° C. Afterloading, the column was washed with additional equilibration buffer andthe protein eluted with equilibration buffer containing 0.25 Mimidazole. The highly purified protein was subsequently desalted into astorage buffer containing 10 mM Hepes, 0.14 M NaCl and 4% mannitol, pH6.8, with a 25 ml G25 Superfine (Pharmacia) column and stored at −80° C.

[2472] Immunoadhesin (Fc containing) constructs of proteins werepurified from the conditioned media as follows. The conditioned mediawere pumped onto a 5 ml Protein A column (Pharmacia) which had beenequilibrated in 20 mM Na phosphate buffer, pH 6.8. After loading, thecolumn was washed extensively with equilibration buffer before elutionwith 100 mM citric acid, pH 3.5. The eluted protein was immediatelyneutralized by collecting 1 ml fractions into tubes containing 275 mL of1 M Tris buffer, pH 9. The highly purified protein was subsequentlydesalted into storage buffer as described above for the poly-His taggedproteins. The homogeneity of the proteins was verified by SDSpolyacrylamide gel (PEG) electrophoresis and N-terminal amino acidsequencing by Edman degradation.

[2473] PRO181, PRO195, PRO200, PRO320, PRO237, PRO273, PRO285, PRO337,PRO526, PRO540, PRO846, PRO362, PRO363, PRO617, PRO322, PRO1083, PRO868,768, PRO792, PRO788, PRO162, PRO1114, PRO827, PRO1075 and PRO1031 weresuccessfully expressed in baculovirus infected Hi5 insect cells. Whilethe expression was actually performed in a 0.5-2 L scale, it can bereadily scaled up for larger (e.g. 8 L) preparations.

[2474] For expression in baculovirus-infected Hi5 insect cells, the PROpolypeptide-encoding DNA may be amplified with suitable systems, such asPfu (Stratagene), or fused upstream (5′-of) of an epitope tag containedwith a baculovirus expression vector. Such epitope tags include poly-histags and immunoglobulin tags (like Fe regions of IgG). A variety ofplasmids may be employed, including plasmids derived from commerciallyavailable plasmids such as pVL1393 (Novagen). Briefly, the PROpolypeptide or the desired portion of the PRO polypeptide (such as thesequence encoding the extracellular domain of a transmembrane protein)is amplified by PCR with primers complementary to the 5′ and 3′ regions.The 5′ primer may incorporate flanking (selected) restriction enzymesites. The product is then digested with those selected restrictionenzymes and subcloned into the expression vector. For example,derivatives of pVL1393 can include the Fc region of human IgG(pb.PH.IgG) or an 8 histidine (pb.PH.His) tag downstream (3′-of) theNAME sequence. Preferably, the vector construct is sequenced forconfirmation.

[2475] Hi5 cells are grown to a confluency of 50% under the conditionsof, 27° C., no CO2, NO pen/strep. For each 150 mm plate, 30 ug of pIEbased vector containing PRO polypeptide is mixed with 1 ml Ex-Cellmedium (Media: Ex-Cell 401+1/100 L-Glu JRH Biosciences #14401-78P (note:this media is light sensitive)), and in a separate tube, 100 ul ofCellfectin (CellFECTIN (GibcoBRL #10362-010) (vortexed to mix)) is mixedwith 1 ml of Ex-Cell medium. The two solutions are combined and allowedto incubate at room temperature for 15 minutes. 8 ml of Ex-Cell media isadded to the 2rnl of DNA/CellFECTIN mix and this is layered on HiS cellsthat have been washed once with Ex-Cell media. The plate is thenincubated in darkness for 1 hour at room temperature. The DNA/CeIIFECTINmix is then aspirated, and the cells are washed once with Ex-Cell toremove excess CellFECTIN . 30 ml of fresh Ex-Cell media is added and thecells are incubated for 3 days at 28° C. The supernatant is harvestedand the expression of the PRO polypeptide in the baculovirus expressionvector can be determined by batch binding of 1 ml of supernatent to 25mL of Ni-NTA beads (QIAGEN) for histidine tagged proteins or Protein-ASepharose CL-4B beads (Pharmacia) for IgG tagged proteins followed bySDS-PAGE analysis comparing to a known concentration of protein standardby Coomassie blue staining.

[2476] The conditioned media from the transfected cells (0.5 to 3 L) isharvested by centrifugation to remove the cells and filtered through0.22 micron filters. For the poly-His tagged constructs, the proteincomprising the PRO polypeptide is purified using a Ni-NTA column(Qiagen). Before purification, imidazole is added to the conditionedmedia to a concentration of 5 mM. The conditioned media is pumped onto a6 ml Ni-NTA column equilibrated in 20 mM Hepes, pH 7.4, buffercontaining 0.3 M NaCl and 5 mM imidazole at a flow rate of 4-5 ml/min.at 4° C. After loading, the column is washed with additionalequilibration buffer and the protein eluted with equilibration buffercontaining 0.25 M imidazole. The highly purified protein is subsequentlydeslated into a storage buffer containing 10 mM Hepes, 0.14 M NaCl and4% mannitol, pH 6.8, with a 25 ml G25 Superfine (Pharmacia) column andstored at −80° C.

[2477] Immunoadhesin (Fc containing) constructs of proteins are purifiedfrom the conditioned media as follows. The conditioned media is pumpedonto a 5 ml Protein A column (Pharmacia) which had been equilibrated in20 mM Na phosphate buffer, pH 6.8. After loading, the column is washedextensively with equilibration buffer before elution with 100 mM citricacid, pH 3.5. The eluted protein is immediately neutralized bycollecting 1 ml fractions into tubes containing 275 mL of 1 M Trisbuffer, pH 9. The highly purified protein is subsequently desalted intostorage buffer as described above for the poly-His tagged proteins. Thehomogeneity of PRO polypeptide can be assessed by SDS polyacrylamidegels and by N-terminal amino acid sequencing by Edman degradation andother analytical procedures as desired or necessary.

[2478] Many of the PRO polypeptides described herein were successfullyexpressed as described above.

Example 104 Preparation of Antibodies that Bind to PRO Polypeptides

[2479] This example illustrates preparation of monoclonal antibodieswhich can specifically bind to a PRO polypeptide.

[2480] Techniques for producing the monoclonal antibodies are known inthe art and are described, for instance, in Goding, supra. Immunogensthat may be employed include purified PRO polypeptide, fusion proteinscontaining the PRO polypeptide, and cells expressing recombinant PROpolypeptide on the cell surface. Selection of the immunogen can be madeby the skilled artisan without undue experimentation.

[2481] Mice, such as Balb/c, are immunized with the PRO polypeptideimmunogen emulsified in complete Freund's adjuvant and injectedsubcutaneously or intraperitoneaUy in an amount from 1-100 micrograms.Alternatively, the immunogen is emulsified in MPL-TDM adjuvant (RibiImmunochemical Research, Hamilton, MT) and injected into the animal'shind foot pads. The immunized mice are then boosted 10 to 12 days laterwith additional immunogen emulsified in the selected adjuvant.Thereafter, for several weeks, the mice may also be boosted withadditional immunization injections. Serum samples may be periodicallyobtained from the mice by retro-orbital bleeding for testing in ELISAassays to detect anti-PRO polypeptide antibodies.

[2482] After a suitable antibody titer has been detected, the animals“positive” for antibodies can be injected with a final intravenousinjection of PRO polypeptide. Three to four days later, the mice aresacrificed and the spleen cells are harvested. The spleen cells are thenfused (using 35% polyethylene glycol) to a selected murine myeloma cellline such as P3X63AgU.1, available from ATCC, No. CRL 1597. The fusionsgenerate hybridoma cells which can then be plated in 96 well tissueculture plates containing HAT (hypoxanthine, aminopterin, and thymidine)medium to inhibit proliferation of non-fused cells, myeloma hybrids, andspleen cell hybrids.

[2483] The hybridoma cells will be screened in an ELISA for reactivityagainst the PRO polypeptide. Determination of “positive” hybridoma cellssecreting the desired monoclonal antibodies against the PRO polypeptideis within the skill in the art.

[2484] The positive hybridoma cells can be injected intraperitoneallyinto syngeneic Balb/c mice to produce ascites containing the anti-PROpolypeptide monoclonal antibodies. Alternatively, the hybridoma cellscan be grown in tissue culture flasks or roller bottles. Purification ofthe monoclonal antibodies produced in the ascites can be accomplishedusing ammonium sulfate precipitation, followed by gel exclusionchromatography. Alternatively, affinity chromatography based uponbinding of antibody to protein A or protein G can be employed.

Example 105 Chimeric PRO Polypeptides

[2485] PRO polypeptides may be expressed as chimeric proteins with oneor more additional polypeptide domains added to facilitate proteinpurification. Such purification facilitating domains include, but arenot limited to, metal chelating peptides such as histidine-tryptophanmodules that allow purification on immobilized metals, protein A domainsthat allow purification on immobilized immunoglobulin, and the domainutilized in the FLAGS™ extension/affimity purification system (hmmunexCorp., Seattle Wash.). The inclusion of a cleavable linker sequence suchas Factor XA or enterokinase (Invitrogen, San Diego Calif.) between thepurification domain and the PRO polypeptide sequence may be useful tofacilitate expression of DNA encoding the PRO polypeptide.

Example 106 Purification of PRO Polypeptides Using Specific Antibodies

[2486] Native or recombinant PRO polypeptides may be purified by avariety of standard techniques in the art of protein purification. Forexample, pro-PRO polypeptide, mature PRO polypeptide, or pre-PROpolypeptide is purified by immunoaffinity chromatography usingantibodies specific for the PRO polypeptide of interest. In general, animmunoaffinity column is constructed by covalently coupling the anti-PROpolypeptide antibody to an activated chromatographic resin.

[2487] Polyclonal immunoglobulins are prepared from immune sera eitherby precipitation with ammonium sulfate or by purification on immobilizedProtein A (Pharmacia LKB Biotechnology, Piscataway, N.J.). Likewise,monoclonal antibodies are prepared from mouse ascites fluid by ammoniumsulfate precipitation or chromatography on immobilized Protein A.Partially purified immunoglobulin is covalently attached to achromatographic resin such as CnBr-activated SEPHAROSE™ (Pharmacia LKBBiotechnology). The antibody is coupled to the resin, the resin isblocked, and the derivative resin is washed according to themnanufacturer's instructions.

[2488] Such an immunoaffinity column is utilized in the purification ofPRO polypeptide by preparing a fraction from cells containing PROpolypeptide in a soluble form. This preparation is derived bysolubilization of the whole cell or of a subcellular fraction obtainedvia differential centrifugation by the addition of detergent or by othermethods well known in the art. Alternatively, soluble PRO polypeptidecontaining a signal sequence may be secreted in useful quantity into themedium in which the cells are grown.

[2489] A soluble PRO polypeptide-containing preparation is passed overthe immunoaffinity column, and the column is washed under conditionsthat allow the preferential absorbance of PRO polypeptide (e.g., highionic strength buffers in the presence of detergent). Then, the columnis eluted under conditions that disrupt antibody/PRO polypeptide binding(e.g., a low pH buffer such as approximately pH 2-3, or a highconcentration of a chaotrope such as urea or thiocyanate ion), and PROpolypeptide is collected.

Example 107 Drug Screening

[2490] This invention is particularly useful for screening compounds byusing PRO polypeptides or binding fragment thereof in any of a varietyof drug screening techniques. The PRO polypeptide or fragment employedin such a test may either be free in solution, affixed to a solidsupport, borne on a cell surface, or located intracellularly. One methodof drug screening utilizes eukaryotic or prokaryotic host cells whichare stably transformed with recombinant nucleic acids expressing the PROpolypeptide or fragment. Drugs are screened against such transformedcells in competitive binding assays. Such cels, either in viable orfixed form, can be used for standard binding assays. One may measure,for example, the formation of complexes between PRO polypeptide or afragment and the agent being tested. Alternatively, one can examine thediminution in complex formation between the PRO polypeptide and itstarget cell or target receptors caused by the agent being tested.

[2491] Thus, the present invention provides methods of screening fordrugs or any other agents which can affect a PRO polypeptide-associateddisease or disorder. These methods comprise contacting such an agentwith an PRO polypeptide or fragment thereof and assaying (I) for thepresence of a complex between the agent and the PRO polypeptide orfragment, or (ii) for the presence of a complex between the PROpolypeptide or fragment and the cell, by methods well known in the art.In such competitive binding assays, the PRO polypeptide or fragment istypically labeled. After suitable incubation, free PRO polypeptide orfragment is separated from that present in bound form, and the amount offree or uncomplexed label is a measure of the ability of the particularagent to bind to PRO polypeptide or to interfere with the PROpolypeptide/cell complex.

[2492] Another technique for drug screening provides high throughputscreening for compounds having suitable binding affinity to apolypeptide and is described in detail in WO 84/03564, published on Sep.13, 1984. Briefly stated, large numbers of different small peptide testcompounds are synthesized on a solid substrate, such as plastic pins orsome other surface. As applied to a PRO polypeptide, the peptide testcompounds are reacted with PRO polypeptide and washed. Bound PROpolypeptide is detected by methods well known in the art. Purified PROpolypeptide can also be coated directly onto plates for use in theaforementioned drug screening techniques. In addition, non-neutralizingantibodies can be used to capture the peptide and immobilize it on thesolid support.

[2493] This invention also contemplates the use of competitive drugscreening assays in which neutralizing antibodies capable of binding PROpolypeptide specifically compete with a test compound for binding to PROpolypeptide or fragments thereof. In this manner, the antibodies can beused to detect the presence of any peptide which shares one or moreantigenic determinants with PRO polypeptide.

Example 108 Rational Drug Design

[2494] The goal of rational drug design is to produce structural analogsof biologically active polypeptide of interest (i.e., a PRO polypeptide)or of small molecules with which they interact, e.g., agonists,antagonists, or inhibitors. Any of these examples can be used to fashiondrugs which are more active or stable forms of the PRO polypeptide orwhich enhance or interfere with the function of the PRO polypeptide invivo (cf., Hodgson, Bio/Technology, 9: 19-21 (1991)).

[2495] In one approach, the three-dimensional structure of the PROpolypeptide, or of an PRO polypeptide-inhibitor complex, is determinedby x-ray crystallography, by computer modeling or, most typically, by acombination of the two approaches. Both the shape and charges of the PROpolypeptide must be ascertained to elucidate the structure and todetermine active site(s) of the molecule. Less often, useful informationregarding the structure of the PRO polypeptide may be gained by modelingbased on the structure of homologous proteins. In both cases, relevantstructural information is used to design analogous PRO polypeptide-likemolecules or to identify efficient inhibitors. Useful examples ofrational drug design may include molecules which have improved activityor stability as shown by Braxton and Wells, Biochemistry, 31:7796-7801(1992) or which act as inhibitors, agonists, or antagonists of nativepeptides as shown by Athauda et al., J. Biochem. 113:742-746 (1993).

[2496] It is also possible to isolate a target-specific antibody,selected by functional assay, as described above, and then to solve itscrystal structure. This approach, in principle, yields a pharmacore uponwhich subsequent drug design can be based. It is possible to bypassprotein crystallography altogether by generating anti-idiotypicantibodies (anti-ids) to a functional, pharmacologically activeantibody. As a mirror image of a mirror image, the binding site of theant-ids would be expected to be an analog of the original receptor. Theanti-id could then be used to identify and isolate peptides from banksof chemically or biologically produced peptides. The isolated peptideswould then act as the pharmacore.

[2497] By virtue of the present invention, sufficient amounts of the PROpolypeptide may be made available to perform such analytical studies asX-ray crystallography. In addition, knowledge of the PRO polypeptideamino acid sequence provided herein will provide guidance to thoseemploying computer modeling techniques in place of or in addition tox-ray crystallography.

Example 109 Ability of PRO Polypeptides to Inhibit Vascular EndothelialGrowth Factor (VEGF) Stimulated Proliferation of Endothelial Cell Growth(Assay 9)

[2498] The ability of various PRO polypeptides to inhibit VEGFstimulated proliferation of endothelial cells was tested. Polypeptidestesting positive in this assay are useful for inhibiting endothelialcell growth in mammals where such an effect would be beneficial, e.g.,for inhibiting tumor growth.

[2499] Specifically, bovine adrenal cortical capillary endothelial cells(ACE) (from primary culture, maximum of 12-14 passages) were plated in96-well plates at 500 cells/well per 100 microliter. Assay mediaincluded low glucose DMEM, 10% calf serum, 2 mM glutamine, and 1×penicillin/streptomycin/fungizone. Control wells included the following:(1) no ACE cells added; (2) ACE cells alone; (3) ACE cells plus 5 ng/mlFGF; (4) ACE cells plus 3 ng/ml VEGF; (5) ACE cells plus 3 ng/ml VEGFplus 1 ng/ml TGF-beta; and (6) ACE cells plus 3 ng/ml VEGF plus 5 ng/mlLIF. The test samples, poly-his tagged PRO polypeptides (in 100microliter volumes), were then added to the wells (at dilutions of 1%,0.1% and 0.01%, respectively). The cell cultures were incubated for &7days at 37° C./5% CO₂. After the incubation, the media in the wells wasaspirated, and the cells were washed 1× with PBS. An acid phosphatasereaction mixture (100 microliter; 0.1M sodium acetate, pH 5.5, 0.1%Triton X-100, 10 mM p-nitrophenyl phosphate) was then added to eachwell. After a 2 hour incubation at 37° C., the reaction was stopped byaddition of 10 microliters 1N NaOH. Optical density (OD) was measured ona microplate reader at 405 nm.

[2500] The activity of PRO polypeptides was calculated as the percentinhibition of VEGF (3 ng/ml) stimulated proliferation (as determined bymeasuring acid phosphatase activity at OD 405 nm) relative to the cellswithout stimulation. TGF-beta was employed as an activity reference at 1nglml, since TGF-beta blocks 70-90% of VEGF-stimulated ACE cellproliferation. The results are indicative of the utility of the PROpolypeptides in cancer therapy and specifically in inhibiting tumorangiogenesis. Numerical values (relative inhibition) are determined bycalculating the percent inhibition of VEGF stimulated proliferation bythe PRO polypeptides relative to cells without stimulation and thendividing that percentage into the percent inhibition obtained by TGF-βat 1 ng/ml which is known to block 70-90% of VEGF stimulated cellproliferation. The results are considered positive if the PROpolypeptide exhibits 30% or greater inhibition of VEGF stimulation ofendothelial cell growth (relative inhibition 30% or greater).

[2501] The following polypeptides tested positive in this assay: PRO200,PRO322 and PRO320.

Example 110 Retinal Neuron Survival (Assay 52)

[2502] This example demonstrates that certain PRO polypeptides haveefficacy in enhancing the survival of retinal neuron cells and,therefore, are useful for the therapeutic treatment of retinal disordersor injuries including, for example, treating sight loss in mammals dueto retinitis pigmentosum, AMD, etc.

[2503] Sprague Dawley rat pups at postnatal day 7 (mixed population:glia and retinal neuronal types) are killed by decapitation followingCO₂ anesthesia and the eyes are removed under sterile conditions. Theneural retina is dissected away from the pigment epithelium and otherocular tissue and then dissociated into a single cell suspension using0.25% trypsin in Ca²⁺, Mg²⁺-free PBS. The retinas are incubated at 37°C. for 7-10 minutes after which the trypsin is inactivated by adding 1ml soybean trypsin inhibitor. The cells are plated at 100,000 cells perwell in 96 well plates in DMEM/F12 supplemented with N2 and with orwithout the specific test PRO polypeptide. Cells for all experiments aregrown at 37° C. in a water saturated atmosphere of 5% CO₂. After 2-3days in culture, cells are stained with calcein AM then fixed using 4%paraformaldehyde and stained with DAPI for determination of total cellcount. The total cells (fluorescent) are quantified at 20× objectivemagnification using CCD camera and NIH image software for MacIntosh.Fields in the well are chosen at random.

[2504] The effect of various concentration of PRO polypeptides arereported herein where percent survival is calculated by dividing thetotal number of calcein AM positive cells at 2-3 days in culture by thetotal number of DAPI-labeled cells at 2-3 days in culture. Anythingabove 30% survival is considered positive.

[2505] The following PRO polypeptides tested positive in this assayusing polypeptide concentrations within the range of 0.01% to 1.0% inthe assay: PRO200, PRO322, PRO540, PRO846 and PRO617.

Example 111 Rod Photoreceptor Survival (Assay 56)

[2506] This assay shows that certain polypeptides of the invention actto enhance the survival/proliferation of rod photoreceptor cells and,therefore, are useful for the therapeutic treatment of retinal disordersor injuries including, for example, treating sight loss in mammals dueto retinitis pigmentosum, AMD, etc. Sprague Dawley rat pups at 7 daypostnatal (mixed population: glia and retinal neuronal cell types) arekilled by decapitation following CO₂ anesthesis and the eyes are removedunder sterile conditions. The neural retina is dissected away form thepigment epithelium and other ocular tissue and then dissociated into asingle cell suspension using 0.25% trypsin in Ca²⁺, Mg²⁺-free PBS. Theretinas are incubated at 37° C. for 7-10 minutes after which the trypsinis inactivated by adding 1 ml soybean trypsin inhibitor. The cells areplated at 100,000 cells per well in 96 well plates in DMEM/F12supplemented with N₂. Cells for all experiments are grown at 37° C. in awater saturated atmosphere of 5% CO₂. After 2-3 days in culture, cellsare fixed using 4% paraformaldehyde, and then stained using CellTrackerGreen CMFDA. Rho 4D2 (ascites or IgG 1:100), a monoclonal antibodydirected towards the visual pigment rhodopsin is used to detect rodphotoreceptor cells by indirect immunofluorescence. The results arecalculated as % survival: total number of calcein—rhodopsin positivecells at 2-3 days in culture, divided by the total number of rhodopsinpositive cells at time 2-3 days in culture. The total cells(fluorescent) are quantified at 20× objective magnification using a CCDcamera and NIH image software for Macintosh. Fields in the well arechosen at random.

[2507] The following polypeptides tested positive in this assay: PRO200,PRO322, PRO540, PRO846 and PRO617.

Example 112 Ability of PRO Polypeptides to Stimulate the Release ofProteoglycans from Cartilage (Assay 97)

[2508] The ability of various PRO polypeptides to stimulate the releaseof proteoglycans from cartilage tissue was tested as follows.

[2509] The metacarphophalangeal joint of 4-6 month old pigs wasaseptically dissected, and articular cartilage was removed by free handslicing being careful to avoid the underlying bone. The cartilage wasminced and cultured in bulk for 24 hours in a humidified atmosphere of95% air, 5% CO₂ in serum free (SF) media (DME/F12 1:1) woth 0.1% BSA and100 U/ml penicillin and 100 mg/ml streptomycin. After washing threetimes, approximately 100 mg of articular cartilage was aliquoted intomicronics tubes and incubated for an additional 24 hours in the above SFmedia. PRO polypeptides were then added at 1% either alone or incombination with 18 ng/ml interleukin-1α, a known stimulator ofproteoglycan release from cartilage tissue. The supernatant was thenharvested and assayed for the amount of proteoglycans using the1,9-dimethyl-methylene blue (DMB) coloriietric assay (Farndale andButtle, Biochem. Biophys. Acta 883:173-177 (1985)). A positive result inthis assay indicates that the test polypeptide will fmd use, forexample, in the treatment of sports-related joint problems, articularcartilage defects, osteoarthritis or rheumatoid arthritis.

[2510] When various PRO polypeptides were tested in the above assay, thepolypeptides demonstrated a marked ability to stimulate release ofproteoglycans from cartilage tissue both basally and after stimulationwith interleulin-1α and at 24 and 72 hours after treatment, therebyindicating that these PRO polypeptides are useful for stimulatingproteoglycan release from cartilage tissue. As such, these PROpolypeptides are useful for the treatment of sports-related jointproblems, articular cartilage defects, osteoarthritis or rheumatoidarthritis. The polypeptides testing positive in this assay are: PRO200.

Example 113 In vitro Antiproliferative Assay (Assay 161)

[2511] The antiproliferative activity of various PRO polypeptides wasdetermined in the investigational, disease-oriented in vitro anti-cancerdrug discovery assay of the National Cancer Institute (NCI), using asulforhodamine B (SRB) dye binding assay essentially as described bySkehan et al., J. Natl. Cancer Inst. 82:1107-1112 (1990). The 60 tumorcell lines employed in this study (“the NCI panel”), as well asconditions for their maintenance and culture in vitro have beendescribed by Monks et al., J. Natl. Cancer Inst. 83:757-766 (1991). Thepurpose of this screen is to initially evaluate the cytotoxic and/orcytostatic activity of the test compounds against different types oftumors (Monks et al., supra; Boyd, Cancer: Princ. Pract. Oncol. Update3(10):1-12 [1989]).

[2512] Cells from approximately 60 human tumor cell lines were harvestedwith trypsin/EDTA (Gibco), washed once, resuspended in IMEM and theirviability was determined. The cell suspensions were added by pipet (100μL volume) into separate 96-well microtiter plates. The cell density forthe 6day incubation was less than for the 2day incubation to preventovergrowth. Inoculates were allowed a preincubation period of 24 hoursat 37° C. for stabilization. Dilutions at twice the intended testconcentration were added at time zero in 100 μL aliquots to themicrotiter plate wells (1:2 dilution). Test compounds were evaluated atfive half-log dilutions (1000 to 100,000-fold). Incubations took placefor two days and six days in a 5% CO₂ atmosphere and 100% humidity.

[2513] After incubation, the medium was removed and the cells were fixedin 0.1 ml of 10% trichloroacetic acid at 40° C. The plates were rinsedfive times with deionized water, dried, stained for 30 minutes with 0.1ml of 0.4% sulforhodamine B dye (Sigma) dissolved in 1% acetic acid,rinsed four times with 1% acetic acid to remove unbound dye, dried, andthe stain was extracted for five minutes with 0.1 ml of 10 mM Tris base[tris(hydroxymethyl)aminomethane], pH 10.5. The absorbance (OD) ofsulforhodamine B at 492 nm was measured using a computer-interfaced,96-well microtiter plate reader.

[2514] A test sample is considered positive if it shows at least 50%growth inhibitory effect at one or more concentrations. PRO polypeptidestesting positive in this assay are shown in Table 7, where theabbreviations are as follows: TABLE 7 Test compound Tumor Cell Line TypeCell Line Designation PRO181 Leukemia RPMI-8226 PRO181 NSCL NCI-H226;NCL-H522 PRO181 Melanoma MALME-3M; SK-MEL-5 PRO181 Ovarian OVCAR-4PRO181 Breast NCI/ADR-RES PRO181 Leukemia MOLT-4 PRO181 NSCL NCI-H226*PRO181 CNS SNB-19 PRO181 Ovarian OVCAR-3; OVCAR-8 PRO181 Renal A498PRO181 Breast MDA-MB-231/ATCC; MDA-N PRO181 Melanoma LOX IMVI PRO181Leukemia CCRF-CEM; RPMI-8226* PRO181 NSCL HOP-62 PRO181 Leukemia HL-60(TB) PRO237 Leukemia K-562 PRO237 NSCL NCI-H322M PRO237 Colon HCC-2998;HCT-15 PRO237 Colon KM12 PRO237 Prostate DU-145 PRO237 Breast MDA-NPRO526 NSCL HOP-62; NCI-H322M PRO526 Colon HCT-116 PRO526 Melanoma LOXLMVI; SK-MEL-2 PRO526 Ovarian OVCAR-3 PRO526 Prostate PC-3 PRO526 NSCLNCI-H226 PRO526 CNS SF-539 PRO526 Renal CAKI-1; RXF 393 PRO362 NSCLNCI-H322M PRO362 Colon HCT-116 PRO362 CNS SF-295 PRO362 Melanoma LOXIMVI PRO362 Leukemia MOLT-4; RPMI-8226; SR PRO362 Colon COLO 205 PRO362Breast HS 578T; MDA-N PRO362 Prostate PC-3 PRO362 Leukemia HL-60 (TB);K-562 PRO362 NSCL EKVX; NCI-H23 PRO362 Colon HCC-2998 PRO362 CNS U251PRO362 Melanoma UACC-257; UACC-62 PRO362 Ovarian OVCAR-8 PRO362 BreastT-47D PRO362 NSCL NCI-H522 PRO362 Renal RXF 393; UO-31 PRO362 BreastMDA-MB-435 PRO362 NSCL HOP-62; NCI-H522 PRO362 Colon KMl2 PRO362Melanoma MALME-3M; SK-MEL-2 PRO362 Melanoma SK-MEL-28; SK-MEL-5 PRO362Ovarian OVCAR-3; OVCAR-4 PRO362 Breast MCF7 PRO866 Leukemia HL-60 (TB);MOLT-4; SR PRO866 NSCL HOP-62 PRO866 NSCL HOP-92 PRO866 Colon KM12PRO866 CNS SF-295 PRO866 Ovarian IGROV1 PRO866 Breast MDA-MB-435 PRO866Melanoma LOX IMVI PRO320 Leukemia CCRF-CEM; RPMI-8226 PRO320 NSCL HOP62;NCI H322M PRO320 Colon HCT-116 PRO320 Renal SN12C PRO320 Breast MDA-NPRO320 Ovarian OVCAR-3 PRO320 Melanoma MALME-3M

[2515] The results of these assays demonstrate that the positive testingPRO polypeptides are useful for inhibiting neoplastic growth in a numberof different tumor cell types and may be used therapeutically therefor.Antibodies against these PRO polypeptides are useful for affinitypurification of these useful polypeptides. Nucleic acids encoding thesePRO polypeptides are useful for the recombinant preparation of thesepolypeptides.

Example 114 Gene Amplification in Tumors

[2516] This example shows that certain PRO polypeptide-encoding genesare amplified in the genome of certain human lung, colon and/or breastcancers and/or cell lines. Amplification is associated withoverexpression of the gene product, indicating that the polypeptides areuseftu targets for therapeutic intervention in certain cancers such ascolon, lung, breast and other cancers and diagnostic determination ofthe presence of those cancers. Therapeutic agents may take the form ofantagonists of the PRO polypeptide, for example, murine-human chimeric,humanized or human antibodies against a PRO polypeptide.

[2517] The starting material for the screen was genomic DNA isolatedfrom a variety cancers. The DNA is quantitated precisely, e.g.,fluorometrically. As a negative control, DNA was isolated from the cellsof ten normal healthy individuals which was pooled and used as assaycontrols for the gene copy in healthy individuals (not shown). The 5′nuclease assay (for example, TaqMan™) and real-time quantitative PCR(for example, ABI Prizm7700 Sequence Detection System™ (Perkin Elmer,Applied Biosystems Division, Foster City, Calif.)), were used to findgenes potentially amplified in certain cancers. The results were used todetermine whether the DNA encoding the PRO polypeptide isover-represented in any of the primary lung or colon cancers or cancercell lines or breast cancer cell lines that were screened. The primarylung cancers were obtained from individuals with tumors of the type andstage as indicated in Table 8. An explanation of the abbreviations usedfor the designation of the primary tumors listed in Table 8 and theprimary tumors and cell lines referred to throughout this example aregiven below.

[2518] The results of the TaqMan™ are reported in delta (Δ) Ct units.One unit corresponds to 1 PCR cycle or approximately a 2-foldamplification relative to normal, two units corresponds to 4-fold, 3units to 8-fold amplification and so on. Quantitation was obtained usingprimers and a TaqMan™ fluorescent probe derived from the PROpolypeptide-encoding gene. Regions of the PRO polypeptide-encoding genewhich are most likely to contain unique nucleic acid sequences and whichare least likely to have spliced out introns are preferred for theprimer and probe derivation, e.g., 3′-untranslated regions. Thesequences for the primers and probes (forward, reverse and probe) usedfor the PRO polypeptide gene amplification analysis were as follows:PRO853 (DNA48227-1350) 48227.tm.f1 5′-GGCACTTCATGGTCCTTGAAA-3′ (SEQ IDNO:539) 48227.tm.p1 5′-CGGATGTGTGTGAGGCCATGCC-3′ (SEQ ID NO:540)48227.tm.r1 5′-GAAAGTAACCACGGAGGTCAAGAT-3′ (SEQ ID NO:541) PRO1017(DNA56112-1379): 56112.tm.f1 5′-CCTCCTCCGAGACTGAAAGCT-3′ (SEQ ID NO:542)56112.tm.p1 5′-TCGCGTTGCTTTTTCTCGCGTG-3′ (SEQ ID NO:543) 56112.tm.r15′-GCGTGCGTCAGGTTCCA-3′ (SEQ ID NO:544) PRO213-1 (DNA30943-1163-1):30943.tm.f3: 5′-CGTTCGTGCAGCGTGTGTA-3′ (SEQ ID NO:545) 30943.tm.p3:5′-CTTCCTCACCACCTGCGACGGG-3′ (SEQ ID NO:546) 30943.tm.r3:5′-GGTAGGCGGTCCTATAGATGGTT-3′ (SEQ ID NO:547) 30943.tm.f1:5′-AGATGTGGATGAATGCAGTGCTA-3′ (SEQ ID NO:548) 30943.tm.p1:5′-ATCAACACCGCCGGCAGTTACTGG-3′ (SEQ ID NO:549) 30943.tm.r1:5′-ACAGAGTGTACCGTCTGCAGACA-3′ (SEQ ID NO:550) 30943.3trn-5:5′-AGCCTCCTGGTGCACTCCT-3′ (SEQ ID NO:551) 30943.3trn-probe:5′-CGACTCCCTGAGCGAGCAGATTTCC-3′ (SEQ ID NO:552) 30943.3trn-3:5′-GCTGGGCAGTCACGAGTCTT-3′ (SEQ ID NO:553) PRO237 (DNA34353-1428):34353.tm.f: 5′-AATCCTCCATCTCAGATCTTCCAG-3′ (SEQ ID NO:554) 34353.tm.p:5′-CCTCAGCGGTAACAGCCGTCC-3′ (SEQ ID NO:555) 34353.tm.r:5′-TGGGCCAAGGGCTGC-3′ (SEQ ID NO:556) PRO324 (DNA36343-1310):36343.tmf1: 5′-TGGTGGATAACCAACAAGATGG-3′ (SEQ ID NO:557) 36343.tmp1:5′-GAGTCTGCATCCACACCACTCTTAAAGTTCTCAA-3′ (SEQ ID NO:558) 36343.tmr1:5′-CAGGTGCTCTTTTCAGTCATGTTT-3′ (SEQ ID NO:559) PRO351 (DNA40571-1315):40571.tm.f1: 5′-TGGCCATTCTCAGGACAAGAG-3′ (SEQ ID NO:560) 40571.tm.p1:5′-CAGTAATGCCATTTGCCTGCCTGCAT-3′ (SEQ ID NO:561) 40571.tm.r1:5′-TGCCTGGAATCACATGACA-3′ (SEQ ID NO:562) PRO362 (DNA45416-1251):45416.tm.f1: 5′-TGTGGCACAGACCCAATCCT-3′ (SEQ ID NO:563) 45416.tm.p1:5′-GACCCTGAAGGCCTCCGGCCT-3′ (SEQ ID NO:564) 45416.tm.r1:5′-GAGAGAGGGAAGGCAGCTATGTC-3′ (SEQ ID NO:565) PRO615 (DNA48304-1323):48304.tm.f1: 5′-CAGCCCCTCTCTTTCACCTGT-3′ (SEQ ID NO:566) 48304.tm.p1:5′-CCATCCTGTGCAGCTGACACACAGC-3′ (SEQ ID NO:567) 48304.tm.r1: 5′-GCCAGGCTATGA GGCTCCTT-3′ (SEQ ID NO:568) PRO531 (DNA48314-1320):48314.tm.f1: 5′-TTCAAGTTCCTGAAGCCGATTAT-3′ (SEQ ID NO:569) 48814.tm.p1:5′-CCAACTTCCCTCCCCAGTGCCCT-3′ (SEQ ID NO:570) 48814.tm.r1:5′-TTGGGGAAGGTAGAATTTCCTTGTAT-3′ (SEQ ID NO:571) PRO618 (DNA49152-1324):49152.tm.f1: 5′-CCCTTCTGCCTCCCAATTCT-3′ (SEQ ID NO:572) 49152.tm.p1:5′-TCTCCTCCGTCCCCTTCCTCCACT-3′ (SEQ ID NO:573) 49152.tm.r1:5′-TGAGCCACTGCCTTGCATTA-3′ (SEQ ID NO:574) PRO772 (DNA49645-1347):49645.tm.f2: 5′-TCTGCAGACGCGATGGATAA-3′ (SEQ ID NO:575) 49645.tm.p2:5′-CCGAAAATAAAACATCGCCCCTTCTGC-3′ (SEQ ID NO:576) 49645.tm.r2:5′-CACGTGGCCTTTCACACTGA-3′ (SEQ ID NO:577) 49645.tm.f1:5′-ACTTGTGACAGCAGTATGCTGTCTT-3′ (SEQ ID NO:578) 49645.tm.p1:5′-AAGCTTCTGTTCAATCCCAGCGGTCC-3′ (SEQ ID NO:579) 49645.tm.r1:5′-ATGCACAGGCTTTTTCTGGTAA-3′ (SEQ ID NO:580) PRO703 (DNA50913-1287):50913.tm.f1: 5′-GCAGGAAACCTTCGAATCTGAG-3′ (SEQ ID NO:581) 50913.tm.p1:5′-ACACCTGAGGCACCTGAGAGAGGAACTCT-3′ (SEQ ID NO:582) 50913.tm.r1:5′-GACAGCCCAGTACACCTGCAA-3′ (SEQ ID NO:583) PRO792 (DNA56352-1358):56352.tm.f1: 5′-GACGGCTGGATCTGTGAGAAA-3′ (SEQ ID NO:584) 56352.tm.p1:5′-CACAACTGCTGACCCCGCCCA-3′ (SEQ ID NO:585) 56352.tm.r1:5′-CCAGGATACGACATGCTGCAA-3′ (SEQ ID NO:586) PRO474 (DNA56045-1380):56045.tm.f1: 5′-AAACTCCAACCTGTATCAGATGCA-3′ (SEQ ID NO:587) 56045.tm.p1:5′-CCCCCAAGCCCTTAGACTCTAAGCC-3′ (SEQ ID NO:588) 56045.tm.r1:5′-GACCCGGCACCTTGCTAAC-3′ (SEQ ID NO:589) PRO274 (DNA39987-1184):39987.tm.f: 5′-GGACGGTCAGTCAGGATGACA-3′ (SEQ ID NO:590) 39987.tm.p:5′-TTCGGCATCATCTCTTCCCTCTCCC-3′ (SEQ ID NO:591) 39987.tm.r:5′-ACAAAAAAAAGGGAACAAAATACGA-3′ (SEQ ID NO:592) PRO381 (DNA44194-1317)44194.tm.f: 5′-CTTTGAATAGAAGACTTCTGGACAATTT-3′ (SEQ ID NO:593)44194.tm.p: 5′-TTGCAACTGGGAATATACCACGACATGAGA-3′ (SEQ ID NO:594)44194.tm.r: 5′-TAGGGTGCTAATTTGTGCTATAACCT-3′ (SEQ ID NO:595)44194.tm.f2: 5′-GGCTCTGAGTCTCTGCTTGA-3′ (SEQ ID NO:596) 44194.tm.p2:5′-TCCAACAACCATTTTCCTCTGGTCC-3′ (SEQ ID NO:597) 44194.tm.r2:5′-AAGCAGTAGCCATTAACAAGTCA-3′ (SEQ ID NO:598) PRO717 (DNA50988-1326):50988.tm.f3: 5′-CAAGCGTCCAGGTTTATTGA-3′ (SEQ ID NO:599) 50988.tm.r3:5′-GACTACAAGGCGCTCAGCTA-3′ (SEQ ID NO:600) 50988.tm.p3:5′-CCGGCTGGGTCTCACTCCTCC-3′ (SEQ ID NO:601) PRO1330 and PRO1449(DNA64907-1163 and DNA64908-1163. respectively): 30943.tm.f3:5′-CGTTCGTGCAGCGTGTGTA-3′ (SEQ ID NO:602) 30943.tm.p3:5′-CTTCCTCACCACCTGCGACG GG-3′ (SEQ ID NO:603) 30943.tm.r3:5′-GGTAGGCGGTCCTATAGATGGTT-3′ (SEQ ID NO:604) 30943.tm.f1: 5′-AGATGTGGATGAATG CAGTGCTA-3′ (SEQ ID NO:605) 30943.tm.p1:5′-ATCAACACCGCCGGCAGTTACTGG-3′ (SEQ ID NO:606) 30943.tm.r1:5′-ACAGAGTGTACCGTCTGCAGACA-3′ (SEQ ID NO:607) 30943.3trn-5:5′-AGCCTCCTGGTGCACTCCT-3′ (SEQ ID NO:608) 30943.3trn-probe:5′-CGACTCCCTGAGCGAGCAGATTTCC-3′ (SEQ ID NO:609) 30943.3trn-3:5′-GCTGGGCAGTCACGAGTCTT-3′ (SEQ ID NO:610)

[2519] The 5′ nuclease assay reaction is a fluorescent PCR-basedtechnique which makes use of the 5′ exonuclease activity of Taq DNApolymerase enzyme to monitor amplification in real time. Twooligonucleotide primers (forward [.f] and reverse [.r]) are used togenerate an amplicon typical of a PCR reaction. A third oligonucleotide,or probe (.p), is designed to detect nucleotide sequence located betweenthe two PCR primers. The probe is non-extendible by Taq DNA polymeraseenzyme, and is labeled with a reporter fluorescent dye and a quencherfluorescent dye. Any laser-induced emission from the reporter dye isquenched by the quenching dye when the two dyes are located closetogether as they are on the probe. During the amplification reaction,the Taq DNA polymerase enzyme cleaves the probe in a template-dependentmanner. The resultant probe fragments disassociate in solution, andsignal from the released reporter dye is free from the quenching effectof the second fluorophore. One molecule of reporter dye is liberated foreach new molecule synthesized, and detection of the unquenched reporterdye provides the basis for quantitative interpretation of the data.

[2520] The 5′ nuclease procedure is run on a real-time quantitative PCRdevice such as the ABI Prism 7700TM Sequence Detection. The systemconsists of a thermocycler, laser, charge-coupled device (CCD) cameraand computer. The system amplifies samples in a 96-well format on athermocycler. During amplification, laser-induced fluorescent signal iscollected in real-time through fiber optics cables for all 96 wells, anddetected at the CCD. The system includes software for running theinstrument and for analyzing the data.

[2521] 5′ Nuclease assay data are initially expressed as Ct, or thethreshold cycle. This is defined as the cycle at which the reportersignal accumulates above the background level of fluorescence. The ΔCtvalues are used as quantitative measurement of the relative number ofstarting copies of a particular target sequence in a nucleic acid samplewhen comparing cancer DNA results to normal human DNA results.

[2522] Table 8 describes the stage, T stage and N stage of variousprimary tumors which were used to screen the PRO polypeptide compoundsof the invention. TABLE 8 Primary Lung and Colon Tumor Profiles PrimaryTumor Stage Stage Other Stage Dukes Stage T Stage N Stage Human lungtumor AdenoCa (SRCC724) [LT1] IIA T1 N1 Human lung tumor SqCCa (SRCC725)[LT1a] IIB T3 N0 Human lung tumor AdenoCa (SRCC726) [LT2] IB T2 N0 Humanlung tumor AdenoCa (SRCC727) [LT3] IIIA T1 N2 Human lung tumor AdenoCa(SRCC728) [LT4] IB T2 N0 Human lung tumor SqCCa (SRCC729) [LT6] IB T2 N0Human lung tumor Aden/SqCCa (SRCC730) [LT7] IA T1 N0 Human lung tumorAdenoCa (SRCC731) [LT9] IB T2 N0 Human lung tumor SqCCa (SRCC732) [LT10]IIB T2 N1 Human lung tumor SqCCa (SRCC733) [LT11] IIA T1 N1 Human lungtumor AdenoCa (SRCC734) [LT12] IV T2 N0 Human lung tumor AdenoSqCCa(SRCC735)[LT13] IB T2 N0 Human lung tumor SqCCa (SRCC736) [LT15] IB T2N0 Human lung tumor SqCCa (SRCC737) [LT16] IB T2 N0 Human lung tumorSqCCa (SRCC738) [LT17] IIB T2 N1 Human lung tumor SqCCa (SRCC739) [LT18]IB T2 N0 Human lung tumor SqCCa (SRCC740) [LT19] IB T2 N0 Human lungtumor LCCa (SRCC741) [LT21] IIB T3 N1 Human lung AdenoCa (SRCC811)[LT22] 1A T1 N0 Human colon AdenoCa (SRCC742) [CT2] M1 D pT4 N0 Humancolon AdenoCa (SRCC743) [CT3] B pT3 N0 Human colon AdenoCa (SRCC744)[CT8] B T3 N0 Human colon AdenoCa (SRCC745) [CT10] A pT2 N0 Human colonAdenoCa (SRCC746) [CT12] MO, R1 B T3 N0 Human colon AdenoCa (SRCC747)[CT14] pMO, RO B pT3 pN0 Human colon AdenoCa (SRCC748) [CT15] M1, R2 DT4 N2 Human colon AdenoCa (SRCC749) [CT16] pMO B pT3 pN0 Human colonAdenoCa (SRCC750) [CT17] C1 pT3 pN1 Human colon AdenoCa (SRCC751) [CT1]MO, R1 B pT3 N0 Human colon AdenoCa (SRCC752) [CT4] B pT3 M0 Human colonAdenoCa (SRCC753) [CT5] G2 C1 pT3 pN0 Human colon AdenoCa (SRCC754)[CT6] pMO, RO B pT3 pN0 Human colon AdenoCa (SRCC755) [CT7] G1 A pT2 pN0Human colon AdenoCa (SRCC756) [CT9] G3 D pT4 pN2 Human colon AdenoCa(SRCC757) [CT11] B T3 N0 Human colon AdenoCa (SRCC758) [CT18] MO, RO BpT3 pN0

[2523] DNA Preparation:

[2524] DNA was prepared from cultured cell lines, primary tumors, normalhuman blood. The isolation was performed using purification kit, bufferset and protease and all from Quiagen, according to the manufacturer'sinstructions and the description below.

[2525] Cell Culture Lysis:

[2526] Cells were washed and trypsinized at a concentration of 7.5×10⁸per tip and pelleted by centrifuging at 1000 rpm for 5 minutes at 4° C.,followed by washing again with ½ volume of PBS recentrifugation. Thepellets were washed a third time, the suspended cells collected andwashed 2× with PBS. The cells were then suspended into 10 ml PBS. BufferCl was equilibrated at 4° C. Qiagen protease #19155 was diluted into6.25 ml cold ddH₂O to a final concentration of 20 mg/ml and equilibratedat 4PC. 10 ml of G2 Buffer was prepared by diluting Qiagen RNAse A stock(100 mg/ml) to a final concentration of 200 μg/ml.

[2527] Buffer C1 (10 ml, 4° C.) and ddH20 (40 ml, 4° C.) were then addedto the 10 ml of cell suspension, mixed by inverting and incubated on icefor 10 minutes. The cell nuclei were pelleted by centrifuging in aBeckman swinging bucket rotor at 2500 rpm at 4PC for 15 minutes. Thesupernatant was discarded and the nuclei were suspended with a vortexinto 2 ml Buffer C1 (at 4° C.) and 6 ml ddH₂O, followed by a second 4°C. centrifugation at 2500 rpm for 15 minutes. The nuclei were thenresuspended into the residual buffer using 200 μl per tip. G2 buffer (10ml) was added to the suspended nuclei while gentle vortexing wasapplied. Upon completion of buffer addition, vigorous vortexing wasapplied for 30 seconds. Quiagen protease (200 μl, prepared as indicatedabove) was added and incubated at 50° C. for 60 minutes. The incubationand centrifugation was repeated until the lysates were clear (e.g.,incubating additional 30-60 minutes, pelleting at 3000× g for 10 min.,4° C.).

[2528] Solid Human Tumor Sample Preparation And Lysis:

[2529] Tumor samples were weighed and placed mto 50 ml conical tubes andheld on ice. Processing was limited to no more than 250 mg tissue perpreparation (1 tip/preparation). The protease solution was freshlyprepared by diluting into 6.25 ml cold ddH₂O to a final concentration of20 mg/ml and stored at 4° C. G2 buffer (20 ml) was prepared by dilutingDNAse A to a final concentration of 200 mg/ml (from 100 mg/ml stock).The tumor tissue was homogenated in 19 ml G2 buffer for 60 seconds usingthe large tip of the polytron in a laminar- flow TC hood in order toavoid inhalation of aerosols, and held at room temperature. Betweensamples, the polytron was cleaned by spinning at 2×30 seconds each in 2LddH₂O, followed by G2 buffer (50 ml). If tissue was still present on thegenerator tip, the apparatus was disassembled and cleaned.

[2530] Quiagen protease (prepared as indicated above, 1.0 ml) was added,followed by vortexing and incubation at 50° C. for 3 hours. Theincubation and centrifugation was repeated until the lysates were clear(e.g., incubating additional 30-60 minutes, pelleting at 3000× g for 10min., 4° C.).

[2531] Human Blood Preparation and Lysis:

[2532] Blood was drawn from healthy volunteers using standard infectiousagent protocols and citrated into 10 ml samples per tip. Quiagenprotease was freshly prepared by dilution into 6.25 ml cold ddH₂O to afinal concentration of 20 mg/ml and stored at 4° C. G2 buffer wasprepared by diluting RNAse A to a final concentration of 200 μg/ml from100 mg/ml stock. The blood (10 ml) was placed into a 50 ml conical tubeand 10 ml C1 buffer and 30 ml ddH₂O (both previously equilibrated to 4°C.) were added, and the components mixed by inverting and held on icefor 10 minutes. The nuclei were pelleted with a Beckman swinging bucketrotor at 2500 rpm, 4° C. for 15 minutes and the supernatant discarded.With a vortex, the nuclei were suspended into 2 ml C1 buffer (4° C.) and6 ml ddH₂O (4° C.). Vortexing was repeated until the pellet was white.The nuclei were then suspended into the residual buffer using a 200 μltip. G2 buffer (10 ml) were added to the suspended nuclei while gentlyvortexing, followed by vigorous vortexing for 30 seconds. Quiagenprotease was added (2001) and incubated at 50° C. for 60 minutes. Theincubation and centrifugation was repeated until the lysates were clear(e.g., incubating additional 30-60 minutes, pelleting at 3000× g for 10min., 4° C.).

[2533] Purification of Cleared Lysates:

[2534] (1) Isolation of Genomic DNA:

[2535] Genomic DNA was equilibrated (1 sample per maxi tip preparation)with 10 ml QBT buffer. QF elution buffer was equilibrated at 50° C. Thesamples were vortexed for 30 seconds, then loaded onto equilibrated tipsand drained by gravity. The tips were washed with 2×15 ml QC buffer. TheDNA was eluted into 30 ml silanized, autoclaved 30 ml Corex tubes with15 nil QF buffer (50° C.). Isopropanol (10.5 ml) was added to eachsample, the tubes covered with parafin and mixed by repeated inversionuntil the DNA precipitated. Samples were pelleted by centrilgation inthe SS-34 rotor at 15,000 rpm for 10 minutes at 4° C. The pelletlocation was marked, the supernatant discarded, and 10 ml 70% ethanol(4° C.) was added. Samples were pelleted again by centrifugation on theSS-34 rotor at 10,000 rpm for 10 minutes at 49° C. The pellet locationwas marked and the supernatant discarded. The tubes were then placed ontheir side in a drying rack and dried 10 minutes at 37° C., taking carenot to overdry the samples.

[2536] After drying, the pellets were dissolved into 1.0 ml TE (pH 8.5)and placed at 50° C. for 1-2 hours. Samples were held overnight at 4PCas dissolution continued. The DNA solution was then transferred to 1.5mnl tubes with a 26 gauge needle on a tuberculin syringe. The transferwas repeated 5× in order to shear the DNA. Samples were then placed at50° C. for 1-2 hours.

[2537] (2) Ouantitation of Genomic DNA and Preparation for GeneAmplification Assay:

[2538] The DNA levels in each tube were quantified by standard A260,A2$0 spectrophotometry on a 1:20 dilution (5 μl DNA+95 μl ddH₂O) usingthe 0.1 ml quartz cuvetts in the Beckman DU640 spectrophotometer.A₂₆₀/A₂₈₀ ratios were in the range of 1.8-1.9. Each DNA samples was thendiluted further to approximately 200 ng/ml in TE (pH 8.5). If theoriginal material was highly concentrated (about 700 ng/lzl), thematerial was placed at 50° C. for several hours until resuspended.

[2539] Fluorometric DNA quantitation was then performed on the dilutedmaterial (20-600 ng/ml) using the manufacturer's guidelines as modifiedbelow. This was accomplished by allowing a Hoeffer DyNA Quant 200fluorometer to warm-up for about 15 minutes. The Hoechst dye workingsolution (#H33258, 10 μl, prepared within 12 hours of use) was dilutedinto 100 ml 1× TNE buffer. A 2 ml cuvette was filled with thefluorometer solution, placed into the machine, and the machine waszeroed. pGEM 3Zf(+) (2 μl, lot #360851026) was added to 2 ml offluorometer solution and calibrated at 200 units. An additional 2 μl ofpGEM 3Zf(+) DNA was then tested and the reading confirmed at 400+/−10units. Each sample was then read at least in triplicate. When 3 sampleswere found to be within 10% of each other, their average was taken andthis value was used as the quantification value.

[2540] The fluorometricly determined concentration was then used todilute each sample to 10 ng/μl in ddH₂O. This was done simultaneously onall template samples for a single TaqMan plate assay, and with enoughmaterial to run 500-1000 assays. The samples were tested in triplicatewith Taqman™ primers and probe both B-actin and GAPDH on a single platewith normal human DNA and no-template controls. The diluted samples wereused provided that the CT value of normal human DNA subtracted from testDNA was +/−1 Ct. The diluted, lot-qualified genomic DNA was stored in1.0 ml aliquots at −80° C. Aliquots which were subsequently to be usedin the gene amplification assay were stored at 4° C. Each 1 ml aliquotis enough for 8-9 plates or 64 tests.

[2541] Gene Amplification Assay:

[2542] The PRO polypeptide compounds of the invention were screened inthe following primary tumors and the resulting ΔCt values greater thanor equal to 1.0 are reported in Table 9 below. TABLE 9 ΔCt values inlung and colon primary tumor and cell line models PRO1330 Tumor or PROPRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO and CellLine 213-1 237 324 351 362 615 531 853 1017 618 772 703 792 474 274 381717 PRO1449 LT-1 1.60 — — — — — — — — — — — — — — — — 1.60 LT-1a 1.241.04 — — — — 1.70 — 1.785 — 1.33 1.22 1.16 1.94 — — — 1.24 1.62 LT2 — —— — 1.39 — — — — — — — — — — — — — LT3 1.51 1.74 — — — 1.31 1.95 — 2.381.03 1.11 1.77 1.10 2.55 — — — 1.51 1.55 1.24 1.52 1.44 LT4 2.26 — — —1.00 — 1.46 — — — — — — — 1.24 — — 2.26 LT6 1.56 1.16 — — — 1.00 2.07 —2.80 — 1.07 1.15 1.81 2.10 — — — 1.56 2.28 LT7 2.45 1.44 — — — 1.09 — —1.12 — — 1.44 — 1.06 — — — 2.45 1.03 LT9 1.24 — — 1.19 — 1.04 1.10 —2.74 1.39 1.62 — 1.99 2.56 — — — 1.24 1.14 1.11 2.59 LT10 — 1.20 — 1.061.69 1.18 1.96 — 3.52 1.29 1.46 1.48 2.00 2.63 — — — — 1.11 1.16 1.292.85 LT11 2.26 — 1.34 1.02 — 1.46 1.79 1.03 1.54 1.84 1.45 1.90 1.201.36 — — — 2.26 2.85 1.72 2.94 1.83 5.21 2.85 2.25 1.27 1.41 2.25 1.791.25 1.79 1.06 LT12 1.86 — 1.92 — — 2.08 1.86 1.18 1.77 — — 1.38 — 1.64— — — 1.86 4.32 1.87 3.02 1.62 5.01 4.32 2.59 1.41 1.82 2.59 1.55 1.501.55 1.25 LT13 1.98 1.05 — 1.23 — 1.39 2.53 1.33 1.55 — 1.18 1.33 1.331.03 — — 7.03 1.98 2.52 1.09 2.06 2.14 1.20 1.00 2.52 2.38 1.03 1.312.03 4.54 2.38 1.14 1.65 LT15 1.40 — — 1.14 — 1.67 2.56 1.28 2.23 — 1.471.45 1.04 1.35 — — 2.71 1.40 1.58 1.47 2.95 2.01 1.44 1.86 1.58 2.691.09 1.31 2.50 4.97 2.69 1.05 1.52 LT16 1.22 1.22 1.63 1.09 — 1.32 —1.33 2.98 — — 1.07 — 4.23 1.00 — 5.48 1.22 2.77 1.38 1.77 1.52 2.77 1.751.17 1.75 LT17 4.58 1.07 1.75 1.46 — 1.66 1.12 1.21 2.90 1.04 1.42 1.241.35 1.40 — — — 4.58 3.73 1.59 1.53 1.62 1.61 1.115 5.45 3.73 5.55 1.215.55 1.50 1.13 LT18 — — — 1.07 — — — — 3.28 — — — — 5.31 1.61 — — — 1.68LT19 1.03 — 1.90 1.33 — 1.59 2.08 — 2.54 — 1.60 1.38 1.62 1.59 — — —1.03 1.22 1.50 2.95 2.98 1.19 1.84 1.22 1.26 1.03 1.21 4.84 1.26 1.48LT21 1.86 — 1.15 1.27 — 1.19 — — 3.14 — — 1.22 — 5.15 — — — 1.86 1.831.09 1.83 3.21 1.06 3.21 LT22 1.61 — — — — — — — — — — — — — — — — 1.61CT2 1.61 — — — — 1.36 2.21 2.4 3.72 — — 2.10 1.46 2.67 — — — 1.61 2.111.25 2.55 2.55 1.65 2.11 1.90 1.48 CT3 — — — — — 1.12 1.50 1.52 3.91 — —1.62 — 2.41 — — — — 1.58 1.02 CT8 2.80 — — — — — 1.15 1.55 2.66 — — 1.06— 2.34 — — — 2.80 1.34 CT10 2.39 — — — — 1.55 1.75 1.97 3.57 — — 1.96 —2.23 — — — 2.39 1.47 1.78 1.21 CT12 3.45 — — — — 1.08 1.93 1.36 3.50 — —1.57 — 2.46 — — — 3.45 1.30 1.08 CT14 3.79 — — — — 1.76 1.47 1.75 3.88 —— 1.19 — 2.83 — — — 3.79 1.02 1.11 1.86 CT15 3.66 — — — — 1.23 2.44 1.753.62 — — 1.70 — 2.89 — — 2.61 3.66 1.33 CT16 2.66 — — — — 1.29 1.95 1.113.12 — — 1.51 — 2.60 — — 2.21 2.66 CT17 3.63 — — — — 1.44 2.19 1.11 3.34— — 1.31 — 2.33 — — 3.31 3.63 CT1 — — — — — — — 1.09 — — — 1.08 — 1.00 —— — — CT4 1.18 — — — — 1.17 — 1.16 1.11 — — 1.63 — 1.13 — — — 1.18 1.07CT5 1.25 — — — — 1.12 1.59 1.95 2.21 — — 1.50 — 1.84 — — — 1.25 1.161.35 2.05 2.11 CT6 1.27 — — — — — — — 1.12 — — 1.38 — 1.24 — — — 1.271.36 CT7 — — — — — — — 1.14 — — — 1.50 — — — — — — CT9 — — — — — — 1.28— 1.29 — — — — — — — — — CT11 — — — — — 1.74 1.49 1.88 1.48 — — 1.99 —2.11 — — — — 1.17 2.13 CT18 — — — — — 1.36 — — — — — 1.15 — 9.66 — — — —Calu-1 1.35 — — — — — — — — — — — — — — — 1.77 1.35 2.95 2.95 H441 2.00— — — — — — 1.71 — — — — — — — — 2.57 2.00 H522 — — — — — — — 1.03 — — —— — — — — 3.78 — H810 2.76 — — — — — — — — — — — — — — — 1.84 2.76 HT291.31 — — — — — — — — — — — — — — — 1.71 1.31 SW403 2.08 — — — — — — — —— — — — — — — 2.09 2.08 LS174T 1.61 — — — — — — — — — — — — — — — 2.901.61 HCT15 1.22 — — — — — — — — — — — — — — — 1.46 1.22 HCC2998 1.73 — —— — — — — — — — — — — — — 1.20 1.73 HF-000643 — — — — — — — — — — — — —— — 4.83 — — HF-000840 — — — — — — — — — — — — — — — 1.08 — — HF-000811— — — — — — — — — — — — — — — 2.09 — — 3.15 HF-001294 — — — — — — — — —— — — — — — 1.14 — — 1.08 HF-001296 — — — — — — — — — — — — — — — 3.18 —— 3.53 HF-001291 — — — — — — — — — — — — — — — 1.17 — — A549 — — — — — —— — — — — — — — — — 1.66 — H460 — — — — — — — — — — — — — — — — 2.50 —SKMES1 — — — — — — — — — — — — — — — — 2.15 — SW620 — — — — — — — — — —— — — — — — 2.36 — Colo320 — — — — — — — — — — — — — — — — 1.99 — 2.73HCT116 — — — — — — — — — — — — — — — — 1.90 — SKCO1 — — — — — — — — — —— — — — — — 3.13 — Colo205 — — — — — — — — — — — — — — — — 1.48 — KM12 —— — — — — — — — — — — — — — — 1.67 —

[2543] Summary

[2544] Because amplification of the various DNA's as described aboveoccurs in various tumors, it is likely associated with tumor formationand/or growth. As a result, antagonists (e.g., antibodies) directedagainst these polypeptides would be expected to be useful in cancertherapy.

Example 115 Induction of c-fos in Endothelial Cells (Assay 34)

[2545] This assay is designed to determine whether PRO polypeptides showthe ability to induce c-fos in endothelial cells. PRO polypeptidestesting positive in this assay would be expected to be useful for thetherapeutic treatment of conditions or disorders where angiogenesiswould be beneficial including, for example, wound healing, and the like(as would agonists of these PRO polypeptides). Antagonists of the PROpolypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of cancerous tumors.

[2546] Human venous umbilical vein endothelial cells (HUVEC, CellSystems) in growth media (50% Ham's P12 w/o GHT: low glucose, and 50%DMEM without glycine: with NaHCO3, 1% glutamine, 10 mM HEPES, 10% PBS,10 ng/ml bFGF) were plated on 96-well microtiter plates at a celldensity of 1×10⁴cells/well. The day after plating, the cells werestarved by removing the growth media and treating the cells with 100μl/well test samples and controls (positive control=growth media;negative control=Protein32 buffer=10 mM HEPES, 140 mM NaCl, 4% (w/v)mannitol, pH 6.8). The cells were incubated for 30 minutes at 37° C., in5% CO₂. The samples were removed, and the first part of the bDNA kitprotocol (Chiron Diagnostics, cat. #6005-037) was followed, where eachcapitalized reagent/buffer listed below was available from the kit.

[2547] Briefly, the amounts of the TM Lysis Buffer and Probes needed forthe tests were calculated based on information provided by themanufacturer. The appropriate amounts of thawed Probes were added to theTM Lysis Buffer. The Capture Hybridization Buffer was warmed to roomtemperature. The bDNA strips were set up in the metal strip holders, and100 μl of Capture Hybridization Buffer was added to each b-DNA wellneeded, followed by incubation for at least 30 minutes. The test plateswith the cells were removed from the incubator, and the media was gentlyremoved using the vacuum manifold. 100 μl of Lysis Hybridization Bufferwith Probes were quickly pipetted into each well of the microtiterplates. The plates were then incubated at 55° C. for 15 minutes. Uponremoval from the incubator, the plates were placed on the vortex mixerwith the microtiter adapter head and vortexed on the #2 setting for oneminute. 80 μl of the lysate was removed and added to the bDNA wellscontaining the Capture Hybridization Buffer, and pipetted up and down tomix. The plates were incubated at 53° C. for at least 16 hours.

[2548] On the next day, the second part of the bDNA kit protocol wasfollowed. Specifically, the plates were removed from the incubator andplaced on the bench to cool for 10 minutes. The volumes of additionsneeded were calculated based upon information provided by themanufacturer. An Amplifier Working Solution was prepared by making a1:100 dilution of the Amplifier Concentrate (20 fm/μl) in ALHybridization Buffer. The hybridization mixture was removed from theplates and washed twice with Wash A. 50 μl of Amplifier Working Solutionwas added to each well and the wells were incubated at 53° C. for 30minutes. The plates were then removed from the incubator and allowed tocool for 10 minutes. The Label Probe Working Solution was prepared bymaking a 1:100 dilution of Label Concentrate (40 pmoles/μl) in ALHybridization Buffer. After the 10-minute cool-down period, theamplifier hybridization mixture was removed and the plates were washedtwice with Wash A. 50 μl of Label Probe Working Solution was added toeach well and the wells were incubated at 53° C. for 15 minutes. Aftercooling for 10 minutes, the Substrate was warmed to room temperature.Upon addition of 3 μl of Substrate Enhancer to each ml of Substrateneeded for the assay, the plates were allowed to cool for 10 minutes,the label hybridization mixture was removed, and the plates were washedtwice with Wash A and three times with Wash D. 50 μl of the SubstrateSolution with Enhancer was added to each well. The plates were incubatedfor 30 minutes at 37° C. and RLU was read in an appropriate luminometer.

[2549] The replicates were averaged and the coefficient of variation wasdetermined. The measure of activity of the fold increase over thenegative control (Protein 32/HEPES buffer described above) value wasindicated by chemiluminescence units (RLU). The results are consideredpositive if the PRO polypeptide exhibits at least a two-fold value overthe negative buffer control. Negative control=1.00 RLU at 1.00%dilution. Positive control=8.39 RLU at 1.00% dilution.

[2550] The following PRO polypeptides tested positive in this assay:PRO938, PRO200, PRO865, PRO788 and PRO1013.

Example 116 Proliferation of Rat Utricular Supporting Cells (Assay 54)

[2551] This assay shows that certain polypeptides of the invention actas potent mitogens for inner ear supporting cells which are auditoryhair cell progenitors and, therefore, are useful for inducing theregeneration of auditory hair cells and treating hearing loss inmammals. The assay is performed as follows. Rat UEC4 utricularepithelial cells are aliquoted into 96 well plates with a density of3000 cells/well in 200 μl of serum-containing medium at 33° C. The cellsare cultured overnight and are then switched to serum-free medium at 37°C. Various dilutions of PRO polypeptides (or nothing for a control) arethen added to the cultures and the cells are incubated for 24 hours.After the 24 hour incubation, ³H-thymidine (1 μCi/well) is added and thecells are then cultured for an additional 24 hours. The cultures arethen washed to remove unincorporated radiolabel, the cells harvested andCpm per well determined. Cpm of at least 30% or greater in the PROpolypeptide treated cultures as compared to the control cultures isconsidered a positive in the assay.

[2552] The following polypeptide tested positive in this assay: PRO337,PRO363 and PRO1012.

Example 117 Detection of PRO Polypeptides that Affect Glucose or FFAUptake by Primary Rat Adipocytes (Assay 94)

[2553] This assay is designed to determine whether PRO polypeptides showthe ability to affect glucose or FFA uptake by adipocyte cells. PROpolypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of disorders where either thestimulation or inhibition of glucose uptake by adipocytes would bebeneficial including, for example, obesity, diabetes or hyper- orhypo-insulinemia.

[2554] In a 96 well format, PRO polypeptides to be assayed are added toprimary rat adipocytes, and allowed to incubate overnight. Samples aretaken at 4 and 16 hours and assayed for glycerol, glucose and FFAuptake. After the 16 hour incubation, insulin is added to the media andallowed to incubate for 4 hours. At this time, a sample is taken andglycerol, glucose and FFA uptake is measured. Media containing insulinwithout the PRO polypeptide is used as a positive reference control. Asthe PRO polypeptide being tested may either stimulate or inhibit glucoseand FFA uptake, results are scored as positive in the assay if greaterthan 1.5 times or less than 0.5 times the insulin control.

[2555] The following PRO polypeptides tested positive as stimulators ofglucose and/or FFA uptake in this assay: PRO181, PRO200, PRO337, PRO362,PRO363, PRO731, PRO534, PRO1114 and PRO1075.

[2556] The following PRO polypeptides tested positive as inhibitors ofglucose and/or FFA uptake in this assay: PRO195, PRO322, PRO862, PRO868,PRO865 and PRO162.

Example 118 Detection of Polypeptides that Affect Glucose and/or FFAUptake in Skeletal Muscle (Assay 106)

[2557] This assay is designed to determine whether PRO polypeptides showthe ability to affect glucose or FFA uptake by skeletal muscle cells.PRO polypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of disorders where either thestimulation or inhibition of glucose uptake by skeletal muscle would bebeneficial including, for example, diabetes or hyper- orhypo-insulinemia.

[2558] In a 96 well format, PRO polypeptides to be assayed are added toprimary rat differentiated skeletal muscle, and allowed to incubateovernight. Then fresh media with the PRO polypeptide and +/− insulin areadded to the wells. The sample media is then monitored to determineglucose and FFA uptake by the skeletal muscle cells. The insulin willstimulate glucose and FFA uptake by the skeletal muscle, and insulin inmedia without the PRO polypeptide is used as a positive control, and alimit for scoring. As the PRO polypeptide being tested may eitherstimulate or inhibit glucose and FFA uptake, results are scored aspositive in the assay if greater than 1.5 times or less than 0.5 timesthe insulin control.

[2559] The following PRO polypeptides tested positive as eitherstimulatorrs or inhibitors of glucose and/or FFA uptake in this assay:PRO181, PRO200, PRO1083, PRO865, PRO162, PRO1008 and PRO1330.

Example 119 Stimulation of Heart Neonatal Hypertrophy (Assay 1)

[2560] This assay is designed to measure the ability of PRO polypeptidesto stimulate hypertrophy of neonatal heart. PRO polypeptides testingpositive in this assay are expected to be useful for the therapeutictreatment of various cardiac insufficiency disorders.

[2561] Cardiac myocytes from 1-day old Harlan Sprague Dawley rats wereobtained. Cells (180 μL at 7.5×10⁴/ml, serum <0.1%, freshly isolated)are added on day 1 to 96-well plates previously coated with DMEM/F12+4%FCS. Test samples containing the test PRO polypeptide or growth mediumonly (hegative control) (20 μl/well) are added directly to the wells onday 1. PGF (20 μl/well) is then added on day 2 at final concentration of10⁻⁶ M. The cells are then stained on day 4 and visually scored on day5, wherein cells showing no increase in size as compared to negativecontrols are scored 0.0, cells showing a small to moderate increase insize as compared to negative controls are scored 1.0 and cells showing alarge increase in size as compared to negative controls are scored 2.0.A positive result in the assay is a score of 1.0 or greater.

[2562] The following polypeptides tested positive in this assay: PRO195,PRO200, PRO526 and PRO792.

Example 120 Enhancement of Heart Neonatal Hypertrophy Induced by F2a(Assay 37)

[2563] This assay is designed to measure the ability of PRO polypeptidesto stimulate hypertrophy of neonatal heart. PRO polypeptides testingpositive in this assay are expected to be useful for the therapeutictreatment of various cardiac insufficiency disorders.

[2564] Cardiac myocytes from l-day old Harlan Sprague Dawley rats wereobtained. Cells (180 μl at 7.5×10⁴/ml, serum <0.1%, freshly isolated)are added on day 1 to 96-well plates previously coated with DMEM/F12+4%FCS. Test samples containing the test PRO polypeptide (20 μl/well) areadded directly to the wells on day 1. PGF (20 μl/well) is then added onday 2 at a final concentration of 10⁻⁶ M. The cells are then stained onday 4 and visually scored on day 5. Visual scores are based on cellsize, wherein cells showing no increase in size as compared to negativecontrols are scored 0.0, cells showing a small to moderate increase insize as compared to negative controls are scored 1.0 and cells showing alarge increase in size as compared to negative controls are scored 2.0.A score of 1.0 or greater is considered positive.

[2565] No PBS is included, since calcium concentration is critical forassay response. Plates are coated with DMEM/F12 plus 4% FCS (200μl/well). Assay media included: DMEM/F12 (with 2.44 gm bicarbonate), 10/2g/ml transferrin, 1 μg/ml insulin, 1 μg/ml aprotinin, 2 mmol/Lglutamine, 100 U/ml penicillin G, 100 μg/ml streptomycin. Protein buffercontaining mannitol (4%) gave a positive signal (score 3.5) at {fraction(1/10)} (0.4%) and {fraction (1/100)} (0.04%), but not at {fraction(1/1000)} (0.004%). Therefore the test sample buffer containing mannitolis not run.

[2566] The following PRO polypeptides tested positive in this assay:PRO195.

Example 121 Guinea Pig Vascular Leak (Assays 32 and 51)

[2567] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to induce vascular permeability.Polypeptides testing positive in this assay are expected to be usefulfor the therapeutic treatment of conditions which would benefit fromenhanced vascular permeability including, for example, conditions whichmay benefit from enhanced local immune system cell infiltration.

[2568] Hairless guinea pigs weighing 350 grams or more were anesthetizedwith Ketamine (75-80 mg/kg) and 5 mg/kg Xylazine intramuscularly. Testsamples containing the PRO polypeptide or a physiological buffer withoutthe test polypeptide are injected into skin on the back of the testanimals with 100 μl per injection site intradermally. There wereapproximately 16-24 injection sites per animal. One ml of Evans blue dye(1% in PBS) is then injected intracardially. Skin vascular permeabilityresponses to the compounds (i.e., blemishes at the injection sites ofinjection) are visually scored by measuring the diameter (in mm) ofblue-colored leaks from the site of injection at 1 and 6 hours postadministration of the test materials. The mm diameter of blueness at thesite of injection is observed and recorded as well as the severity ofthe vascular leakage. Blemishes of at least 5 mm in diameter areconsidered positive for the assay when testing purified proteins, beingindicative of the ability to induce vascular leakage or permeability. Aresponse greater than 7 mm diameter is considered positive forconditioned media samples. Human VEGF at 0.1 μl/100 μl is used as apositive control, inducing a response of 15-23 mm diameter.

[2569] The following PRO polypeptides tested positive in this assay:PRO200.

Example 122 Skin Vascular Permeability Assay (Assay 64)

[2570] This assay shows that certain polypeptides of the inventionstimulate an immune response and induce inflammation by inducingmononuclear cell, eosinophil and PMN infiltration at the site ofinjection of the animal. Compounds which stimulate an immune responseare useful therapeutically where stimulation of an immune response isbeneficial. This skin vascular permeability assay is conducted asfollows. Hairless guinea pigs weighing 350 grams or more areanesthetized with ketamine (75-80 mg/Kg) and 5 mg/Kg xylazineintramuscularly (IM). A sample of purified polypeptide of the inventionor a conditioned media test sample is injected intradermally onto thebacks of the test animals with 100 μl per injection site. It is possibleto have about 10-30, preferably about 16-24, injection sites per animal.One μl of Evans blue dye (1% in physiologic buffered saline) is injectedintracardially. Blemishes at the injection sites are then measured (mmdiameter) at 1 hr and 6 hr post injection. Animals were sacrificed at 6hrs after injection. Each skin injection site is biopsied and fixed informalin. The skins are then prepared for histopathologic evaluation.Each site is evaluated for inflammatory cell infiltration into the skin.Sites with visible inflammatory cell inflammation are scored aspositive. Inflammatory cells may be neutrophilic, eosinophilic,monocytic or lymphocytic. At least a minimal perivascular infiltrate atthe injection site is scored as positive, no infiltrate at the site ofinjection is scored as negative.

[2571] The following polypeptide tested positive in this assay: PRO200,PRO362 and PRO1031.

Example 123 Induction of c-fos in Cortical Neurons (Assay 83)

[2572] This assay is designed to determine whether PRO polypeptides showthe ability to induce c-fos in cortical neurons. PRO polypeptidestesting positive in this assay would be expected to be useful for thetherapeutic treatment of nervous system disorders and injuries whereneuronal proliferation would be beneficial.

[2573] Cortical neurons are dissociated and plated in growth medium at10,000 cells per well in 96 well plates. After aproximately 2 cellulardivisions, the cells are treated for 30 minutes with the PRO polypeptideor nothing (negative control). The cells are then fixed for 5 minuteswith cold methanol and stained with an antibody directed againstphosphorylated CREB. mRNA levels are then calculated usingchemiluminescence. A positive in the assay is any factor that results inat least a 2-fold increase in c-fos message as compared to the negativecontrols.

[2574] The following PRO polypeptides tested positive in this assay:PRO200.

Example 124 Mouse Kidney Mesangial Cell Proliferation Assay (Assay 92)

[2575] This assay shows that certain polypeptides of the invention actto induce proliferation of mammalian kidney mesangial cells and,therefore, are useful for treating kidney disorders associated withdecreased mesangial cell function such as Berger disease or othernephropathies associated with Schönlein-Henoch purpura, celiac disease,dermatitis herpetiformis or Crohn disease. The assay is performed asfollows. On day one, mouse kidney mesangial cells are plated on a 96well plate in growth media (3:1 mixture of Dulbecco's modified Eagle'smedium and Ham's F12 medium, 95% fetal bovine serum, 5% supplementedwith 14 mM HEPES) and grown overnight. On day 2, PRO polypeptides arediluted at 2 concentrations (1% and 0.1%) in serum-free medium and addedto the cells. Control samples are serum-free medium alone. On day 4, 20μl of the Cell Titer 96 Aqueous one solution reagent (Progema) was addedto each well and the colormetric reaction was allowed to proceed for 2hours. The absorbance (OD) is then measured at 490 nm. A positive in theassay is anything that gives an absorbance reading which is at least 15%above the control reading.

[2576] The following polypeptide tested positive in this assay: PRO200,PRO363, PRO731, PRO534, PRO866 and PRO1031.

Example 125 Pericyte c-Fos Induction (Assay 93)

[2577] This assay shows that certain polypeptides of the invention actto induce the expression of c-fos in pericyte cells and, therefore, areuseful not only as diagnostic markers for particular types ofpericyte-associated tumors but also for giving rise to antagonists whichwould be expected to be useful for the therapeutic treatment ofpericyte-associated tumors. Specifically, on day 1, pericytes arereceived from VEC Technologies and all but 5 ml of media is removed fromflask. On day 2, the pericytes are trypsinized, washed, spun and thenplated onto 96 well plates. On day 7, the media is removed and thepericytes are treated with 100 μl of PRO polypeptide test samples andcontrols (positive control=DME+5% serum +/−PDGF at 500 ng/ml; negativecontrol=protein 32). Replicates are averaged and SD/CV are determined.Fold increase over Protein 32 (buffer control) value indicated bychemiluminescence units (RLU) luminometer reading verses frequency isplotted on a histogram. Two-fold above Protein 32 value is consideredpositive for the assay. ASY Matrix: Growth media low glucose DMEM 32 20%FBS+1× pen strep+1× fungizone. Assay Media=low glucose DMEM+5% FBS.

[2578] The following polypeptides tested positive in this assay: PRO200.

Example 126 Chondrocyte Re-differentiation Assay (Assay 110)

[2579] This assay shows that certain polypeptides of the invention actto induce redifferentiation of chondrocytes, therefore, are expected tobe useful for the treatment of various bone and/or cartilage disorderssuch as, for example, sports injuries and arthritis. The assay isperformed as follows. Porcine chondrocytes are isolated by overnightcollagenase digestion of articulary cartilage of metacarpophalangealjoints of 46 month old female pigs. The isolated cells are then seededat 25,000 cells/cm² in Ham F-12 containing 10% FBS and 4 g/mlgentamycin. The culture media is changed every third day and the cellsare then seeded in 96 well plates at 5,000 cells/well in 100 μl of thesame media without serum and 100 μl of the test PRO polypeptide, 5 nMstaurosporin (positive control) or medium alone (negative control) isadded to give a final volume of 200 μl/well. After 5 days of incubationat 37° C., a picture of each well is taken and the differentiation stateof the chondrocytes is determined. A positive result in the assay occurswhen the redifferentiation of the chondrocytes is determined to be moresimilar to the positive control than the negative control.

[2580] The following polypeptide tested positive in this assay: PRO200,PRO285, PRO337, PRO526, PRO362, PRO363, PRO531, PRO1083, PRO862, PRO733,PRO1017, PRO792, PRO788, PRO1008, PRO1075, PRO725 and PRO1031.

Example 127 Fetal Hemoglobin Induction in an Erythroblastic Cell Line(Assay 107)

[2581] This assay is useful for screening PRO polypeptides for theability to induce the switch from adult hemoglobin to fetal hemoglobinin an erythroblastic cell line. Molecules testing positive in this assayare expected to be useful for therapeutically treating various mammalianhemoglobin-associated disorders such as the various thalassemias. Theassay is performed as follows. Erythroblastic cells are plated instandard growth medium at 1000 cells/well in a 96 well format. PROpolypeptides are added to the growth medium at a concentration of 0.2%or 2% and the cells are incubated for 5 days at 37° C. As a positivecontrol, cells are treated with 100 μM hemin and as a negative control,the cells are untreated. After 5 days, cell lysates are prepared andanalyzed for the expression of gamma globin (a fetal marker). A positivein the assay is a gamma globin level at least 2-fold above the negativecontrol.

[2582] The following polypeptides tested positive in this assay: PRO237,PRO381, PRO362, PRO724, PRO866, PRO1114, PRO725 and PRO1071.

Example 128 Induction of Pancreatic β-Cell Precursor Proliferation(Assay 117)

[2583] This assay shows that certain polypeptides of the invention actto induce an increase in the number of pancreatic β-Cell precursor cellsand, therefore, are useful for treating various insulin deficient statesin mammals, including diabetes mellitus. The assay is performed asfollows. The assay uses a primary culture of mouse fetal pancreaticcells and the primary readout is an alteration in the expression ofmarkers that represent either β-cell precursors or mature β-cells.Marker expression is measured by real time quantitative PCR (RTQ-PCR);wherein the marker being evaluated is a transcription factor calledPdx1.

[2584] The pancreata are dissected from E14 embryos (CDI mice). Thepancreata are then digested with collagenase/dispase in F12/DMEM at 37°C. for 40 to 60 minutes (collagenase/dispase, 1.37 mg/ml, BoehringerMannheim, #1097113). The digestion is then neutralized with an equalvolume of 5% BSA and the cells are washed once with RPMI1640. At day 1,the cells are seeded into 12-well tissue culture plates (precoated withlaminin, 20 μg/ml in PBS, Boehringer Mannheim, #124317). Cells frompancreata from 1-2 embryos are distributed per well. The culture mediumfor this primary cuture is 14F/1640. At day 2, the media is removed andthe attached cells washed with RPMI/1640. Two mls of minimal media areadded in addition to the protein to be tested. At day 4, the media isremoved and RNA prepared from the cells and marker expression analyzedby real time quantitative RT-PCR. A protein is considered to be activein the assay if it increases the expression of the relevant β-cellmarker as compared to untreated controls. 14F/1640 is RPM11640 (Gibco)plus the following:

[2585] group A 1:1000

[2586] group B 1:1000

[2587] recombinant human insulin 10 μg/ml

[2588] Aprotinin (50 μg/ml) 1:2000 (Boehringer manhein #981532)

[2589] Bovine pituitary extract (BPE) 60 μg/ml

[2590] Gentamycin 100 ng/ml

[2591] Group A: (in 10 ml PBS)

[2592] Transferrin, 100 mg (Sigma T2252)

[2593] Epidermal Growth Factor, 100 μg (BRL 100004)

[2594] Triiodothyronine, 10 μl of 5×10⁻⁶ M (Sigma T5516)

[2595] Ethanolamine, 100 μl of 10⁻¹ M (Sigma E0135)

[2596] Phosphoethalamrine, 100 μl of 10⁻¹ M (Sigma P0503)

[2597] Selenium, 4 μl of 10⁻¹ M (Aesar #12574)

[2598] Group C: (in 10 ml 100% ethanol)

[2599] Hydrocortisone, 2 μl of 5×10⁻³ M (Sigma #H0135)

[2600] Progesterone, 100 μl of 1×10⁻³ M (Sigma #P6149)

[2601] Forskolin, 500 μl of 20 mM (Calbiochem #344270)

[2602] Minimal media:

[2603] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/rml),gentamycin (100 ng/ml), aprotinin (50 μg/ml) and BPE (15 μg/ml).

[2604] Defined media:

[2605] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml) and aprotinin (50 μg/ml).

[2606] The following polypeptides tested positive in this assay: PRO237and PRO731.

Example 129 Stimulatory Activity in Mixed Lymphocyte Reaction (MLR)Assay (Assay 24)

[2607] This example shows that certain polypeptides of the invention areactive as a stimulator of the proliferation of stimulated T-lymphocytes.Compounds which stimulate proliferation of lymphocytes are usefiltherapeutically where enhancement of an immune response is beneficial. Atherapeutic agent may take the form of antagonists of the polypeptide ofthe invention, for example, murine-human chimeric, humanized or humanantibodies against the polypeptide.

[2608] The basic protocol for this assay is described in CurrentProtocols in Immunology, unit 3.12; edited by J E Coligan, A MKruisbeek, D H Marglies, E M Shevach, W Strober, National Insitutes ofHealth, Published by John Wiley & Sons, Inc.

[2609] More specifically, in one assay variant, peripheral bloodmononuclear cells (PBMC) are isolated from mammnalian individuals, forexample a human volunteer, by leukopheresis (one donor will supplystimulator PBMCs, the other donor will supply responder PBMCs). Ifdesired, the cells are frozen in fetal bovine serum and DMSO afterisolation. Frozen cells may be thawed overnight in assay media (37° C.,5% CO₂) and then washed and resuspended to 3×10⁶ cells/ml of assay media(RPMI; 10% fetal bovine serum, 1% penicillin/streptomycin, 1% glutamine,1% HEPES, 1% non-essential amino acids, 1% pyruvate). The stimulatorPBMCs are prepared by irradiating the cells (about 3000 Rads).

[2610] The assay is prepared by plating in triplicate wells a mixtureof:

[2611] 100:1 of test sample diluted to 1% or to 0.1%,

[2612] 50:1 of irradiated stimulator cells, and

[2613] 50:1 of responder PBMC cells.

[2614] 100 microliters of cell culture media or 100 microliter ofCD4-IgG is used as the control. The wells are then incubated at 37° C.,5% CO₂ for 4 days. On day 5, each well is pulsed with tritiatedthymidine (1.0 mC/well; Amersham). After 6 hours the cells are washed 3times and then the uptake of the label is evaluated.

[2615] In another variant of this assay, PBMCs are isolated from thespleens of Balblc mice and C57B6 mice. The cells are teased from freshlyharvested spleens in assay media (RPMI; 10% fetal bovine serum, 1%penicillin/streptomycin, 1% glutamine, 1% HEPES, 1% nonessential aminoacids, 1% pyruvate) and the PBMCs are isolated by overlaying these cellsover Lympholyte M (Organon Teknika), centrifuging at 2000 rpm for 20minutes, collecting and washing the mononuclear cell layer in assaymedia and resuspending the cells to 1×10⁷ cells/ml of assay media. Theassay is then conducted as described above.

[2616] Positive increases over control are considered positive withincreases of greater than or equal to 180% being preferred. However, anyvalue greater than control indicates a stimulatory effect for the testprotein.

[2617] The following PRO polypeptides tested positive in this assay:PRO273, PRO526, PRO381, PRO719, PRO866 and PRO1031.

Example 130 Inhibitory Activity in Mixed Lymphocyte Reaction (MLR) Assay(Assay 67)

[2618] This example shows that one or more of the polypeptides of theinvention are active as inhibitors of the proliferation of stimulatedT-lymphocytes. Compounds which inhibit proliferation of lymphocytes areuseful therapeutically where suppression of an immune response isbeneficial.

[2619] The basic protocol for this assay is described in CurrentProtocols in Immunology, unit 3.12; edited by J E Coligan, A MKruisbeek, D H Marglies, E M Shevach, W Strober, National Insitutes ofHealth, Published by John Wiley & Sons, Inc.

[2620] More specifically, in one assay variant, peripheral bloodmononuclear cells (PBMC) are isolated from mmallan individuals, forexample a human volunteer, by leukopheresis (one donor will supplystimulator PBMCs, the other donor will supply responder PBMCs). Ifdesired, the cells are frozen in fetal bovine serum and DMSO afterisolation. Frozen cells may be thawed overnight in assay media (37° C.,5% CO2) and then washed and resuspended to 3×10⁶ cells/ml of assay media(RPMI; 10% fetal bovine serum, 1% penicillinistreptomycin, 1% glutamine,1% HEPES, 1% non-essential amino acids, 1% pyruvate). The stimulatorPBMCs are prepared by irradiating the cells (about 3000 Rads).

[2621] The assay is prepared by plating in triplicate wells a mixtureof:

[2622] 100:1 of test sample diluted to 1% or to 0.1%,

[2623] 50:1 of irradiated stimulator cells, and

[2624] 50:1 of responder PBMC cells.

[2625] 100 microliters of cell culture media or 100 microliter ofCD4-IgG is used as the control. The wells are then incubated at 37° C.,5% CO₂ for 4 days. On day 5, each well is pulsed with tritiatedthymidine (1.0 mC/well; Amersham). After 6 hours the cells are washed 3times and then the uptake of the label is evaluated.

[2626] In another variant of this assay, PBMCs are isolated from thespleens of Balb/c mice and C57B6 mice. The cells are teased from freshlyharvested spleens in assay media (RPMI; 10% fetal bovine serum, 1%penicillin/streptomycin, 1% glutamnine, 1% HEPES, 1% non-essential aminoacids, 1% pyruvate) and the PBMCs are isolated by overlaying these cellsover Lympholyte M (Organon Tekika), centrifuging at 2000 rpm for 20minutes, collecting and washing the mononuclear cell layer in assaymedia and resuspending the cells to 1×10⁷ cells/ml of assay media. Theassay is then conducted as described above.

[2627] Any decreases below control is considered to be a positive resultfor an inhibitory compound, with decreases of less than or equal to 80%being preferred. However, any value less than control indicates aninhibitory effect for the test protein.

[2628] The following polypeptide tested positive in this assay: PRO273,PRO526, PRO381, PRO701l, PRO363, PRO531, PRO1083, PRO865, PRO788 andPRO1114.

Example 131 Fibroblast (BHK-21) Proliferation (Assay 98)

[2629] This assay shows that certain PRO polypeptides of the inventionact to induce proliferation of mammalian fibroblast cells in cultureand, therefore, function as useful growth factors in mammalian systems.The assay is performed as follows. BHK-21 fibroblast cells plated instandard growth medium at 2500 cells/well in a total volume of 100 μl.The PRO polypeptide, β-FGF (positive control) or nothing (negativecontrol) are then added to the wells in the presence of lug/ml ofheparin for a total final volume of 200 μl. The cells are then incubatedat 37° C. for 6 to 7 days. After incubation, the media is removed, thecells are washed with PBS and then an acid phosphatase substratereaction mixture (100 μl/well) is added. The cells are then incubated at37° C. for 2 hours. 10 μl per well of 1N NaOH is then added to stop theacid phosphatase reaction. The plates are then read at OD 405 mn. Apositive in the assay is acid phosphatase activity which is at least 50%above the negative control.

[2630] The following PRO polypeptide tested positive in this assay:PRO273 and PRO731.

Example 132 Induction of Endothelial Cell Apootosis (ELISA) (Assay 109)

[2631] The ability of PRO polypeptides to induce apoptosis inendothelial cells was tested in human venous umbilical vein endothelialcells (HUVEC, Cell Systems) using a 96-well format, in 0% serum mediasupplemented with 100 ng/ml VEGF, 0.1% BSA, 1× penn/strep. A positiveresult in this assay indicates the usefulness of the polypeptide fortherapeutically treating any of a variety of conditions associated withundesired endothelial cell growth including, for example, the inhibitionof tumor growth. The 96-well plates used were manufactured by Falcon(No. 3072). Coating of 96 well plates were prepared by allowinggelatinization to occur for >30 minutes with 100 μl of 0.2% gelatin inPBS solution. The gelatin mix was aspirated thoroughly before platingHUEC cells at a final concentration of 2×10⁴ cells/mi in 10% serumcontaining medium—100 μl volume per well. The cells were grown for 24hours before adding test samples containing the PRO polypeptide ofinterest.

[2632] To all wells, 100 μl of 0% serum media (Cell Systems)complemented with 100 ng/ml VEGF, 0.1% BSA, 1× penn/strep was added.Test samples containing PRO polypeptides were added in triplicate atdilutions of 1%, 0.33% and 0.11%. Wells without cells were used as ablank and wells with cells only were used as a negative control. As apositive control, 1:3 serial dilutions of 50 μl of a 3× stock ofstaurosporine were used. The cells were incubated for 24 to 35 hoursprior to ELISA.

[2633] ELISA was used to determine levels of apoptosis preparingsolutions according to the Boehringer Manual [Boehringer, Cell DeathDetection ELISA plus, Cat No. 1 920 6851. Sample preparations: 96 wellplates were spun down at 1 krpm for 10 minutes (200 g); the supernatantwas removed by fast inversion, placing the plate upside down on a papertowel to remove residual liquid. To each well, 200 μl of 1× Lysis bufferwas added and incubation allowed at room temperature for 30 minuteswithout shaking. The plates were spun down for 10 minutes at 1 krpm, and20 μl of the lysate (cytoplasmic fraction) was transferred intostreptavidin coated MTP. 80 μl of immunoreagent mix was added to the 20μl lystate in each well. The MTP was covered with adhesive foil andincubated at room tempearature for 2 hours by placing it on an orbitalshaker (200 rpm). After two hours, the supernatant was removed bysuction and the wells rinsed three times with 250 μl of 1× incubationbuffer per well (removed by suction). Substrate solution was added (100μl) into each well and incubated on an orbital shaker at roomtemperature at 250 rpm until color development was sufficient for aphotometric analysis (approx. after 10-20 minutes). A 96 well reader wasused to read the plates at 405 nm, reference wavelength, 492 nm. Thelevels obtained for PIN 32 (control buffer) was set to 100%. Sampleswith levels >130% were considered positive for induction of apoptosis.

[2634] The following PRO polypeptides tested positive in this assay:PRO846.

Example 133 Induction of Endothelial Cell Apoptosis (Assay 73)

[2635] The ability of PRO polypeptides to induce apoptosis inendothelial cells was tested in human venous umbilical vein endothelialcells (HUVEC, Cell Systems). A positive test in the assay is indicativeof the usefulness of the polypeptide in therapeutically treating tumorsas well as vascular disorders where inducing apoptosis of endothelialcells would be beneficial.

[2636] The cells were plated on 96-well microtiter plates (Amersham LifeScience, cytostar-T scintillating microplate, RPNQ160, sterile,tissue-culture treated, individually wrapped), in 10% serum (CSG-medium,Cell Systems), at a density of 2×10⁴ cells per well in a total volume of100 μl. On day 2, test samples containing the PRO polypeptide were addedin triplicate at dilutions of 1%, 0.33% and 0.11%. Wells without cellswere used as a blank and wells with cells only were used as a negativecontrol. As a positive control 1:3 serial dilutions of 50 μl of a 3×stock of staurosporine were used. The ability of the PRO polypeptide toinduce apoptosis was determined by processing of the 96 well plates fordetection of Annexin V, a member of the calcium and phospholipid bindingproteins, to detect apoptosis. 0.2 ml Annexin V—Biotin stock solution(100 jg/ml) was diluted in 4.6 ml 2 × Ca²⁺ binding buffer and 2.5% BSA(1:25 dilution). 50 μl of the diluted Annexin V—Biotin solution wasadded to each well (except controls) to a final concentration of 1.0μg/ml. The samples were incubated for 10-15 minutes with Annexin-Biotinprior to direct addition of ³⁵S-Streptavidin. ³⁵S-Streptavidin wasdiluted in 2× Ca²⁺ Binding buffer, 2.5% BSA and was added to all wellsat a final concentration of 3×10⁴ cpm/well. The plates were then sealed,centrifuged at 1000 rpm for 15 minutes and placed on orbital shaker for2 hours. The analysis was performed on a 1450 Microbeta Trilux (Wallac).Percent above background represents the percentage amount of counts perminute above the negative controls. Percents greater than or equal to30% above background are considered positive.

[2637] The following PRO polypeptides tested positive in this assay:PRO719.

Example 134 Human Venous Endothelial Cell Calcium Flux Assay (Assay 68)

[2638] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to stimulate calcium flux inhuman umbilical vein endothelial cells (HUVEC, Cell Systems). Calciuminflux is a well documented response upon binding of certain ligands totheir receptors. A test compound that results in a positive response inthe present calcium influx assay can be said to bind to a specificreceptor and activate a biological signaling pathway in humanendothelial cells. This could ultimately lead, for example, toendothelial cell division, inhibition of endothelial cell proliferation,endothelial tube formation, cell migration, apoptosis, etc.

[2639] Human venous umbilical vein endothelial cells (HUVEC, CellSystems) in growth media (50:50 without glycine, 1% glutamine, 10 mMHepes, 10% FBS, 10 ng/ml bFGF), were plated on 96-well microtiterViewPlates-96 (Packard Instrument Company Part #6005182) microtiterplates at a cell density of 2×10⁴ cells/well. The day after plating, thecells were washed three times with buffer (HBSS plus 10 mM Hepes),leaving 100 μl/well. Then 100 μl/well of 8 μM Fluo-3 (2×) was added. Thecells were incubated for 1.5 hours at 37° C./5% CO₂. After incubation,the cells were then washed 3× with buffer (described above) leaving 100μI/well. Test samples of the PRO polypeptides were prepared on different96-well plates at 5× concentration in buffer. The positive controlcorresponded to 50 μM ionomycin (5×); the negative control correspondedto Protein 32. Cell plate and sample plates were run on a FLWPR(Molecular Devices) machine. The FLIPR machine added 25 μl of testsample to the cells, and readings were taken every second for oneminute, then every 3 seconds for the next three minutes.

[2640] The fluorescence change from baseline to the maximum rise of thecurve (A change) was calculated, and replicates averaged. The rate offluorescence increase was monitored, and only those samples which had aA change greater than 1000 and a rise within 60 seconds, were consideredpositive.

[2641] The following PRO polypeptides tested positive in the presentassay: PRO771.

Example 135 Induction of c-fos in Endothelial Cells (Assay 34)

[2642] This assay is designed to determine whether PRO polypeptides showthe ability to induce c-fos in endothelial cells. PRO polypeptidestesting positive in this assay would be expected to be useful for thetherapeutic treatment of conditions or disorders where angiogenesiswould be beneficial including, for example, wound healing, and the like(as would agonists of these PRO polypeptides). Antagonists of the PROpolypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of cancerous tumors.

[2643] Human venous umbilical vein endothelial cells (ATVEC, CellSystems) in growth media (50% Ham's F12 w/o GHT: low glucose, and 50%DMEM without glycine: with NaHCO3, 1% glutamine, 10 mM HEPES, 10% FBS,10 ng/ml bFGF) were plated on 96-well microtiter plates at a celldensity of 1×10⁴ cells/well. The day after plating, the cells werestarved by removing the growth media and treating the cells with 100μl/well test samples and controls (positive control=growth media;negative control=Protein32 buffer=10 mM HEPES, 140 mM NaCl, 4% (w/v)mannitol, pH 6.8). The cells were incubated for 30 minutes at 37° C., in5% CO₂. The samples were removed, and the first part of the bDNA kitprotocol (Chiron Diagnostics, cat. #6005-037) was followed, where eachcapitalized reagent/buffer listed below was available from the kit.

[2644] Briefly, the amounts of the TM Lysis Buffer and Probes needed forthe tests were calculated based on information provided by themanufacturer. The appropriate amounts of thawed Probes were added to theTM Lysis Buffer. The Capture Hybridization Buffer was warmed to roomtemperature. The bDNA strips were set up in the metal strip holders, and100 μl of Capture Hybridization Buffer was added to each b-DNA wellneeded, followed by incubation for at least 30 minutes. The test plateswith the cells were removed from the incubator, and the media was gentlyremoved using the vacuum manifold. 100 μl of Lysis Hybridization Bufferwith Probes were quickly pipetted into each well of the microtiterplates. The plates were then incubated at 55° C. for 15 minutes. Uponremoval from the incubator, the plates were placed on the vortex mixerwith the microtiter adapter head and vortexed on the #2 setting for oneminute. 80 μl of the lysate was removed and added to the bDNA wellscontaining the Capture Hybridization Buffer, and pipetted up and down tomix. The plates were incubated at 53° C. for at least 16 hours.

[2645] On the next day, the second part of the bDNA kit protocol wasfollowed. Specifically, the plates were removed from the incubator andplaced on the bench to cool for 10 minutes. The volumes of additionsneeded were calculated based upon information provided by themanufacturer. An Amplifier Working Solution was prepared by making a1:100 dilution of the Amplifier Concentrate (20 fm/ill) in ALHybridization Buffer. The hybridization mixture was removed from theplates and washed twice with Wash A. 50 ll of Amplifier Working Solutionwas added to each well and the wells were incubated at 53° C. for 30minutes. The plates were then removed from the incubator and allowed tocool for 10 minutes. The Label Probe Working Solution was prepared bymaking a 1:100 dilution of Label Concentrate (40 pmoles/μl) in ALHybridization Buffer. After the 10-minute cool-down period, theamplifier hybridization mixture was removed and the plates were washedtwice with Wash A. 50 μl of Label Probe Working Solution was added toeach well and the wells were incubated at 53° C. for 15 minutes. Aftercooling for 10 minutes, the Substrate was warmed to room temperature.Upon addition of 3 μl of Substrate Enhancer to each ml of Substrateneeded for the assay, the plates were allowed to cool for 10 minutes,the label hybridization mixture was removed, and the plates were washedtwice with Wash A and three times with Wash D. 50 μl of the SubstrateSolution with Enhancer was added to each well. The plates were incubatedfor 30 minutes at 37° C. and RLU was read in an appropriate luminometer.

[2646] The replicates were averaged and the coefficient of variation wasdetermined. The measure of activity of the fold increase over thenegative control (Protein 32/HEPES buffer described above) value wasindicated by chemiluminescence units (RLU). The results are consideredpositive if the PRO polypeptide exhibits at least a two-fold value overthe negative buffer control. Negative control=1.00 RLU at 1.00%dilution. Positive control 8.39 RLU at 1.00% dilution.

[2647] The following PRO polypeptides tested positive in this assay:PRO474.

Example 136 Induction of Pancreatic β-Cell Precursor Differentiation(Assay 89)

[2648] This assay shows that certain polypeptides of the invention actto induce differentiation of pancreatic β-cell precursor cells intomature pancreatic β-cells and, therefore, are useful for treatingvarious insulin deficient states in mammals, including diabetesmellitus. The assay is performed as follows. The assay uses a primaryculture of mouse fetal pancreatic cells and the primary readout is analteration in the expression of markers that represent either β-cellprecursors or mature β-cells. Marker expression is measured by real timequantitative PCR (RTQ-PCR); wherein the marker being evaluated isinsulin.

[2649] The pancreata are dissected from E14 embryos (CD1 mice). Thepancreata are then digested with collagenase/dispase in F12/DMEM at 37°C. for 40 to 60 minutes (collagenase/dispase, 1.37 mg/ml, BoehringerMannheim, #1097113). The digestion is then neutralized with an equalvolume of 5% BSA and the cells are washed once with RPMI1640. At day 1,the cells are seeded into 12-well tissue culture plates (precated withlaminin, 20 μg/ml in PBS, Boehringer Mannheim, #124317). Cells frompancreata from 1-2 embryos are distributed per well. The culture mediumfor this primary cuture is 14F/1640. At day 2, the media is removed andthe attached cells washed with RPMI/1640. Two mls of minimal media areadded in addition to the protein to be tested. At day 4, the media isremoved and RNA prepared from the cells and marker expression analyzedby real time quantitative RT-PCR. A protein is considered to be activein the assay if it increases the expression of the relevant β-cellmarker as compared to untreated controls. 14F/1640 is RPMI1640 (Gibco)plus the following:

[2650] group A 1:1000

[2651] group B 1:1000

[2652] recombinant human insulin 10 μg/ml

[2653] Aprotinin (50 μg/ml) 1:2000 (Boehringer manheim #981532)

[2654] Bovine pituitary extract (13PE) 60 μg/ml

[2655] Gentamycin 100 ng/ml

[2656] Group A: (in 10 ml PBS)

[2657] Transferrin, 100 mg (Sigma T2252)

[2658] Epidermal Growth Factor, 100 μg (BRL 100004)

[2659] Triiodothyronine, 10 μl of 5×10⁴ M (Sigma T5516)

[2660] Ethanolamine, 100 μl of 10⁻¹ M (Sigma E0135)

[2661] Phosphoethalamine, 100 μl of 10⁻¹ M (Sigma P0503)

[2662] Selenium, 4 μl of 10⁻¹ M (Aesar #12574)

[2663] Group C: (in 10 ml 100% ethanol)

[2664] Hydrocortisone, 2 μl of 5×10⁻³ M (Sigma #H0135)

[2665] Progesterone, 100 μl of 1×10⁻³ M (Sigma #P6149)

[2666] Forskolin, 500 μl of 20 mM (Calbiochem #344270)

[2667] Minimal media:

[2668] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml), aprotinin (50 μg/ml) and BPE (15 μg/ml).

[2669] Defined media:

[2670] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml) and aprotinin (50 μg/ml).

[2671] The following polypeptides were positive in this assay: PRO788and PRO162.

Example 137 Stimulation of Endothelial Cell Proliferation (Assay 8)

[2672] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to stimulate adrenal corticalcapillary endothelial cell (ACE) growth. PRO polypeptides testingpositive in this assay would be expected to be useful for thetherapeutic treatment of conditions or disorders where angiogenesiswould be beneficial including, for example, wound healing, and the like(as would agonists of these PRO polypeptides). Antagonists of the PROpolypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of cancerous tumors.

[2673] Bovine adrenal cortical capillary endothelial (ACE) cells (fromprimary culture, maximum of 12-14 passages) were plated in 96-wellplates at 500 cells/well per 100 microliter. Assay media included lowglucose DMEM, 10% calf serum, 2 mM glutamine, and 1×penicillin/streptomycin/fungizone. Control wells included the following:(1) no ACE cells added; (2) ACE cells alone; (3) ACE cells plus VEGF (5ng/ml); and (4) ACE cells plus FGF (5ng/ml). The control or test sample,(in 100 microliter volumes), was then added to the wells (at dilutionsof 1%, 0.1% and 0.01%, respectively). The cell cultures were incubatedfor 6-7 days at 37° C./5% CO₂. After the incubation, the media in thewells was aspirated, and the cells were washed 1× with PBS. An acidphosphatase reaction mixture (100 microliter; 0. IM sodium acetate, pH5.5, 0.1% Triton X-100, 10 mM p-nitrophenyl phosphate) was then added toeach well. After a 2 hour incubation at 37° C., the reaction was stoppedby addition of 10 microliters 1N NaOH. Optical density (OD) was measuredon a microplate reader at 405 nm.

[2674] The activity of a PRO polypeptide was calculated as the foldincrease in proliferation (as determined by the acid phosphataseactivity, OD 405 um) relative to (1) cell only background, and (2)relative to maximum stimulation by VEGF. VEGF (at 3-10 ng/ml) and FGF(at 1-5 ng/nm) were employed as an activity reference for maximumstimulation. Results of the assay were considered “positive” if theobserved stimulation was 2 50% increase over background. VEGF (5 ng/ml)control at 1% dilution gave 1.24 fold stimulation; FGF (5 ng/ml) controlat 1% dilution gave 1.46 fold stimulation.

[2675] The following PRO polypeptides tested positive in this assay:PRO1075.

Example 138 Mouse Mesengial Cell Inhibition Assay (Assay 114)

[2676] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to inhibit the proliferation ofmouse mesengial cells in culture. PRO polypeptides testing positive inthis assay would be expected to be useful for the therapeutic treatmentof such diseases or conditions where inhibition of mesengial cellproliferation would be beneficial such as, for example, cystic renaldysplasia, polycystic kidney disease, or other kidney diseaseassoiciated with abnormal mesengial cell proliferation, renal tumors,and the like.

[2677] On day 1, mouse mesengial cells are plated on a 96 well plate ingrowth medium (a 3:1 mixture of Dulbecco's modified Eagle's medium andHam's F12medium, 95%; fetal bovine serum, 5%; supplemented with 14 mMHEPES) and then are allowed to grow overnight. On day 2, the PROpolypeptide is diluted at 2 different concentrations (1%, 0.1%) inserum-free medium and is added to the cells. The negative control isgrowth medium without added PRO polypeptide. After the cells are allowedto incubate for 48 hours, 20 μl of the Cell Titer 96 Aqueous onesolution reagent (Promega) is added to each well and the colormetricreaction is allowed to proceed for 2 hours. The absorbance (OD) is thenmeasured at 490 nm. A positive in the assay is an absorbance readingwhich is at least 10% above the negative control.

[2678] The following PRO polypeptides tested positive in this assay:PRO200 and PRO697.

Example 139 Chondrocyte Proliferation Assay (Assay 111)

[2679] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to induce the proliferationand/or redifferentiation of chondrocytes in culture. PRO polypeptidestesting positive in this assay would be expected to be useful for thetherapeutic treatment of various bone and/or cartilage disorders suchas, for example, sports injuries and arthritis.

[2680] Porcine chondrocytes are isolated by overnight collagenasedigestion of articular cartilage of the metacarpophalangeal joint of 46month old female pigs. The isolated cells are then seeded at 25,000cells/cm² in Ham F-12 containing 10% FBS and 4 μg/ml gentamycin. Theculture media is changed every third day and the cells are reseeded to25,000 cells/cm² every five days. On day 1-2, the cells are seeded in 96well plates at 5,000 cells/well in 100 μl of the same media withoutserum and 100 μl of either serum-free medium (negative control),staurosporin (final concentration of 5 nM; positive control) or the testPRO polypeptide are added to give a final volume of 200 μl/well. After 5days at 37° C., 20 μl of Alamar blue is added to each well and theplates are incubated for an additional 3 hours at 37° C. Thefluorescence is then measured in each well (Ex: 530 nm; Em: 590 um). Thefluorescence of a plate containing 200 μl of the serum-free medium ismeasured to obtain the background. A positive result in the assay isobtained when the fluorescence of the PRO polypeptide treated sample ismore like that of the positive control than the negative control.

[2681] The following PRO polypeptides tested positive in this assay:PRO181, PRO200 and PRO322.

Example 140 Rat DRG Neuronal Survival Inhibition Assay

[2682] This assay is designed to determine whether PRO polypeptides ofthe present invention show the ability to inhibit the survival of neuralcells in culture. Polypeptides testing positive in this assay areexpected to be useful for the therapeutic treatment of neuropathicconditions which are associated with undesirable neural cellproliferation including, for example, neuroblastomas, gliomas,glioblastomas, and the like.

[2683] A heterogeneous population of neural cells freshly isolated fromE14 rat embryo dorsal root ganglia are diluted in complete medium andare plated at 5,000 cells/well on polyurethane pretreated platescontaining 50 μl F12 complete media. Test PRO polypeptides (50 μl, oneconcentration) with 50 μl additional assay media are then added to testfor survival inhibition activity. Negative controls are treated with 100μl of complete medium alone. After 3 days incubation, the cells arestained with CMFDA and fixed after 1 hour with 4% paraformaldehyde.Cells are then quantified by NIH image analysis. A positive in the assayis cell numbers in the treated well(s) being less than 0.5 of theuntreated control well(s).

[2684] The following PRO polypeptides tested positive in this assay:PRO195 and PRO701.

Example 141 Tissue Expression Distribution

[2685] Oligonucleotide probes were constructed from some of the PROpolypeptide-encoding nucleotide sequences shown in the accompanyingfigures for use in quantitative PCR amplification reactions. Theoligonucleotide probes were chosen so as to give an approximately200-600 base pair amplified fragment from the 3′ end of its associatedtemplate in a standard PCR reaction. The oligonucleotide probes wereemployed in standard quantitative PCR amplification reactions with cDNAlibraries isolated from different human adult and/or fetal tissuesources and analyzed by agarose gel electrophoresis so as to obtain aquantitative determination of the level of expression of the PROpolypeptide-encoding nucleic acid in the various tissues tested.Knowledge of the expression pattern or the differential expression ofthe PRO polypeptide-encoding nucleic acid in various different humantissue types provides a diagnostic marker useful for tissue typing, withor without other tissue- specific markers, for determining the primarytissue source of a metastatic tumor, and the like. These assays providedthe following results. Tissues With Tissues Lacking DNA MoleculeSignificant Expression Significant Expression DNA40954-1233 liver, lungbrain DNA41404-1352 lung, kidney liver, retina, pancreas DNA44179-1362liver lung, brain DNA45234-1277 kidney liver, placenta, brainDNA45415-1318 thyroid, brain, kidney liver, bone marrow DNA45417-1432thyroid, brain, kidney, liver bone marrow DNA45493-1349 liver, kidneybrain DNA48306-1291 brain, kidney pancreas, liver DNA48328-1355 thyroid,brain, bone marrow liver, kidney DNA48329-1290 brain, bone marrow,liver, thyroid kidney DNA49624-1279 placenta liver, lung, kidney, brainDNA50911-1288 brain placenta DNA50914-1289 brain, kidney, liver placentaDNA53906-1368 lung, kidney brain DNA53912-1457 lung, liver, kidney,brain pancreas DNA53977-1371 lung, liver, kidney, brain, pancreas bonemarrow DNA54002-1367 bone marrow, liver, lung, thyroid, brain kidneyDNA55737-1345 bone marrow, kidney liver, brain DNA57039-1402 pigmentepithelium lung, brain, liver, kidney DNA57253-1382 lung, brain, liver,placenta kidney DNA58747-1384 lung, brain, kidney, pancreas, thyroidliver DNA23318-1211 spleen, brain, heart, cartilage colon tumor,prostate DNA39975-1210 brain, colon tumor, THP-1 macrophages heartDNA39979-1213 dendrocytes, cartilage, spleen, substantia heart nigra,uterus, prostate DNA41386-1316 HUVEC, cartilage, substantia nigra, colontumor, dendrocytes uterus DNA50919-1361 HUVEC, brain, prostate,cartilage, spleen, colon tumor heart, uterus DNA52185-1370 dendrocytessubstantia nigra, hippocampus, uterus DNA42663-1154 uterus, spleen,cartilage, HUVEC, bone marrow colon tumor DNA50980-1286 placenta,adrenal bone marrow, gland, prostate uterus, cartilage

Example 142 In situ Hybridization

[2686] In situ hybridization is a powerful and versatile technique forthe detection and localization of nucleic acid sequences within cell ortissue preparations. It may be useful, for example, to identify sites ofgene expression, analyze the tissue distribution of transcription,identify and localize viral infection, follow changes in specific mRNAsynthesis and aid in chromosome mapping.

[2687] In situ hybridization was performed following an optimizedversion of the protocol by Lu and Gillett, Cell Vision 1:169-176 (1994),using PCR-generated ³³P-labeled riboprobes. Briefly, formalin-fixed,paraffin-embedded human tissues were sectioned, deparaffinized,deproteinated in proteinase K (20 g/ml) for 15 minutes at 37° C., andfurther processed for in situ hybridization as described by Lu andGillett, supra. A [³³-P] UTP-labeled antisense riboprobe was generatedfrom a PCR product and hybridized at 55° C. overnight. The slides weredipped in Kodak NTB2 nuclear track emulsion and exposed for 4 weeks.

[2688]³³P-Riboprobe Synthesis

[2689] 6.0 μl (125 mCi) of ³³P-UTP (Amersham BF 1002, SA<2000 Ci/mmol)were speed vac dried. To each tube containing dried ³³P-UTP, thefollowing ingredients were added:

[2690] 2.0 μl 5× transcription buffer

[2691] 1.0 μl DTT (100 mM)

[2692] 2.0 μl NTP mix (2.5 mM: 10μ; each of 10 mM GTP, CTP & ATP+10 μlH₂O)

[2693] 1.0 μl UTP (50 μM)

[2694] 1.0 μl Rnasin

[2695] 1.0 μl DNA template (1 μg)

[2696] 1.0 μl H₂O

[2697] 1.0 μl RNA polymerase (for PCR products T3=AS, T7=S, usually)

[2698] The tubes were incubated at 37° C. for one hour. 1.0 μl RQ1 DNasewere added, followed by incubation at 37° C. for 15 minutes. 90 μl TE(10 mM Tris pH 7.6/1 mM EDTA pH 8.0) were added, and the mixture waspipetted onto DE81 paper. The remaining solution was loaded in aMicrocon-50 ultrafiltration unit, and spun using program 10 (6 minutes).The filtration unit was inverted over a second tube and spun usingprogram 2 (3 minutes). After the final recovery spin, 100 μl TE wereadded. 1 μl of the final product was pipetted on DE81 paper and countedin 6 ml of Biofluor II.

[2699] The probe was run on a TBE/urea gel. 1-3 μl of the probe or 5 μlof RNA Mrk III were added to 3 μl of loading buffer. After heating on a95° C. heat block for three minutes, the gel was immediately placed onice. The wells of gel were flushed, the sample loaded, and run at180-250 volts for 45 minutes. The gel was wrapped in saran wrap andexposed to XAR film with an intensifying screen in −70° C. freezer onehour to overnight.

[2700]³³P-Hybridization

[2701] A. Pretreatment of Frozen Sections

[2702] The slides were removed from the freezer, placed on aluminiumtrays and thawed at room temperature for 5 minutes. The trays wereplaced in 55° C. incubator for five minutes to reduce condensation. Theslides were fixed for 10 minutes in 4% paraformaldehyde on ice in thefume hood, and washed in 0.5× SSC for 5 minutes, at room temperature (25ml 20× SSC+975 ml SQ H₂O). After deproteination in 0.5 μg/ml proteinaseK for 10 minutes at 37° C. (12.5 μl of 10 mg/mn stock in 250 mlprewarmed RNase-free RNAse buffer), the sections were washed in 0.5× SSCfor 10 minutes at room temperature. The sections were dehydrated in 70%,95%, 100% ethanol, 2 minutes each.

[2703] B. Pretreatment of Paraffin-Embedded Sections

[2704] The slides were deparaffinized, placed in SQ H₂O, and rinsedtwice in 2× SSC at room temperature, for 5 minutes each time. Thesections were deproteinated in 20 μg/ml proteinase K (500 μl of 10 mg/mlin 250 ml RNase-free RNase buffer; 37° C., 15 minutes)—human embryo, or8× proteinase K (100 μl in 250 ml Rnase buffer, 37° C., 30 minutes)-formalin tissues. Subsequent rinsing in 0.5× SSC and dehydration wereperformed as described above.

[2705] C. Prehybridization

[2706] The slides were laid out in a plastic box lined with Box buffer(4× SSC, 50% formamide)- saturated filter paper. The tissue was coveredwith 50 μl of hybridization buffer (3.75 g Dextran Sulfate+6 ml SQ H₂O),vortexed and heated in the microwave for 2 minutes with the caploosened. After cooling on ice, 18.75 ml formamide, 3.75 ml 20× SSC and9 ml SQ H₂O were added, the tissue was vortexed well, and incubated at42° C. for 1-4 hours.

[2707] D. Hybridization

[2708] 1.0×10⁶ cpm probe and 1.0 μl tRNA (50 mg/ml stock) per slide wereheated at 95° C. for 3 minutes. The slides were cooled on ice, and 48 μlhybridization buffer were added per slide. After vortexing, 50 μl ³³Pmix were added to 50 μl prehybridization on slide. The slides wereincubated overnight at 55° C.

[2709] E. Washes

[2710] Washing was done 2×10 minutes with 2× SSC, EDTA at roomtemperature (400 ml 20× SSC+16 ml 0.25 M EDTA, V_(f)=4L), followed byRNaseA treatment at 37° C. for 30 minutes (500 μl of 10 mg/ml in 250 mlRnase buffer=20 μg/ml), The slides were washed 2×10 minutes with 2× SSC,EDTA at room temperature. The stringency wash conditions were asfollows: 2 hours at 55° C., 0.1× SSC, EDTA (20 ml 20× SSC+16 ml EDTA,V_(f)=4L).

[2711] F. Oligonucleotides

[2712] In situ analysis was performed on a variety of DNA sequencesdisclosed herein. The oligonucleotides employed for these analyses werederived from the nucleotide sequences disclosed herein and generallyrange from about 40 to 55 nucleotides in length.

[2713] G. Results

[2714] In situ analysis was performed on a variety of DNA sequencesdisclosed herein. The results from these analyses are as follows.

[2715] (1) DNA29101-1122 (PRO200)

[2716] Fetal:

[2717] Lower limb expression in developing lower limb bones at the edgeof the cartilagenous anlage (i.e. around the outside edge); indeveloping tendons, in vascular smooth muscle and in cells embracingdeveloping skeletal muscle myocytes and myotubes. Expression alsoobserved at the epiphyseal growth plate. Lymph node expression inmarginal sinus of developing lymph nodes. Thymus expression in thesubcapsular region of the thymic cortex, possibly representing eitherthe subcapsular epithelial cells or the proliferating, double negative,thymocytes that are found in this region. Spleen is negative. Tracheaexpression in smooth muscle. Brain (cerebral cortex) focal expression incortical neurones. Spinal cord negative. Small intestine expression insmooth muscle. Thyroid—generalized expression over thyroid epithelium.Adrenal is negative. Liver expression in ductal plate cells. Stomachexpression in mural smooth muscle. Fetal skin expression in basal layerof squamous epithelium. Placenta expression in interstitial cells introphoblastic villi. Cord expression in wall of arteries and vein.

[2718] Comments:

[2719] Expression pattern suggests that PRO200 may be involved in celldifferentiation/proliferation.

[2720] High expression was observed at the following additional sites:Chimp ovary—granulosa cells of maturing follicles, lower intensitysignal observed over the cal cells. Chimp parathyroid—high expressionover chief cells. Human fetal testis—moderate expression over stromalcells surrounding developing tubules. Human fetal lung—high expressionover chondrocytes in developing bronchial tree, and low level expressionover branching bronchial epithelium. Specific expression was notobserved over the renal cell, gastric and colonic carcinomas. Fetaltissues examined (E12-E16 weeks) include: placenta, umbilical cord,liver, kidney, adrenals, thyroid, lungs, heart, great vessels,oesophagus, stomach, small intestine, spleen, thymus, pancreas, brain,eye, spinal cord, body wall, pelvis and lower limb. Adult tissuesexamined: liver, kidney, adrenal, myocardium, aorta, spleen, lymph node,pancreas, lung, skin, cerebral cortex (rm), hippocampus(rm),cerebellum(rm), penis, eye, bladder, stomach, gastric carcinoma, colon,colonic carcinoma and chondrosarcoma. Acetominophen induced liver injuryand hepatic cirrhosis.

[2721] (2) DNA30867-1335 (PRO218)

[2722] Low level expression over numerous epithelia including fetalsmall intestine, fetal thyroid, chimp gastric epithelium. Expressionalso seen over malignant cells in a renal cell carcinoma. Expression infetal brain, over cortex. The distribution does not suggest an obviousfunction. Human fetal tissues examined (E12-E16 weeks) include:placenta, umbilical cord, liver, kidney, adrenals, thyroid, lungs,heart, great vessels, oesophagus, stomach, small intestine, spleen,thymus, pancreas, brain, eye, spinal cord, body wall, pelvis and lowerlimb. Adult human tissues examined: kidney (normal and end-stage),bladder, adrenal, spleen, lymph node, pancreas, lung, skin, eye (inc.retina), colon, bladder, liver (normal, cirrhotic, acute failure),heart, clear cell carcinoma of kidney, gastric adenocarcinoma,colorectal carcinoma. Non-human primate tissues examined: Chimp tissues:salivary gland, stomach, thyroid, parathyroid, tongue, thymus, ovary,lymph node, peripheral nerve. Rhesus Monkey tissues: cerebral cortex,hippocampus, cerebellum, penis.

[2723] (3) DNA40021-1154 (PRO285)

[2724] Low levels of expression observed in the placenta and overhematopoietic cells in the mouse fetal liver. No expression was detectedin either human fetal, adult or chimp lymph node and no expression wasdetected in human fetal or human adult spleen. Fetal tissues examined(E12-E16 weeks) include: placenta, umbilical cord, liver, kidney,adrenals, thyroid, lungs, heart, great vessels, oesophagus, stomach,small intestine, spleen, thymus, pancreas, brain, eye, spinal cord, bodywall, pelvis and lower limb. Adult tissues examined: liver, kidney,adrenal, myocardium, aorta, spleen, lymph node, pancreas, lung, skin,cerebral cortex (rm), hippocampus(rm), cerebellum(rm), brain infarct(human), cerebritis (human),penis, eye, bladder, stomach, gastriccarcinoma, colon, colonic carcinoma, thyroid (chimp), parathyroid(chimp) ovary (chimp) and chondrosarcoma. Acetominophen induced liverinjury and hepatic cirrhosis.

[2725] (4) DNA39523-1192 (PRO273)

[2726] Expression over epithelium of mouse embryo skin as well as overbasal epithelium and dermis of human fetal skin. Basal epithelial pegsof the squamous mucosa of the chimp tongue are also positive. Expressionover a subset of cells in developing glomeruli of fetal kidney, adultrenal tubules, and over “thyroidized” epithelium in end-stage renaldisease, low expression in a renal cell carcinoma, probably over theepithelial cells. Low level expression over stromal cells in fetal lung.Expression over stromal cells in the apical portion of gastric glands.High expression in the lamina propria of the fetal small intestinalvilli, normal colonic mucosa and over stromal cells in a coloniccarcinoma. Strong expression over benign connective tissue cells in thehylanized stroma of a sarcoma. Expression over stromal cells in theplacental villi and the splenic red pulp. In the brain, expression overcortical neurones. Connective tissue surrounding developing bones andover nerve sheath cells in the fetus. Fetal tissues examined (E12-E16weeks) include: placenta, umbilical cord, liver, kidney, adrenals,thyroid, lungs, heart, great vessels, oesophagus, stomach, smallintestine, spleen, thymus, pancreas, brain, eye, spinal cord, body wall,pelvis and lower limb. Adult tissues examined: liver, kidney, adrenal,myocardium, aorta, spleen, lymph node, pancreas, lung, skin, cerebralcortex (rm), hippocampus(rm), eye, stomach, gastric carcinoma, colon,colonic carcinoma, thyroid (chimp), parathyroid (chimp) ovary (chimp)and chondrosarcoma. Acetominophen induced liver injury and hepaticcirrhosis.

[2727] Expression was present in many cells in the outer layers (and II)of the monkey cerebral cortex. A small subset of cells in the deepercortical layers also expressed mRNA for this chemokine homolog.Scattered cells within the molecular layers of the hippocampus andbordering the inner edge of the dentate gyrus contained chemokinehomolog mRNA. No expression was detected within the cerebellar cortex.Chemokine homolog expression is not observed in infarcted brain, wherecell death has occurred in the regions where the chemokine homolognormally is expressed. This probe could possibly serve as a marker of asubset of neurons of outer layers of the cerebral cortex and couldpossibly reveal neuronal migration disorders. Abnormal neuronalmigration is a possible cause of some seizure disorders andschizophrenia. In order to gain a better appreciation of thedistribution of this mRNA we will test whether the probe willcross-hybridize with mouse brain tissue.

[2728] Also shows intriguing and specific patterns of hybridizationwithin postnatal day (P) 10 and adult mouse brains. In one sagittalsection of P10 mouse brain, strong signal was observed scatered withinthe molecular layer of the hippocampus and inner edges of the dentategyrus. Cells in the presubiculum were moderately labeled; the signalextended in a strong band through outer layers of the retrosplenialcortes to the occipital cortex, where the signal diminished tobackground levels. A small set of positive neurons were detected indeeper regions of P10 motor cortex; neurons in outer layers of P10cortex did not exhibit signal above background levels. Moderatehybridization signal was also detected in the inferior colliculus.Chemokine homolog signal in the adult mouse brain was evaluated in threecoronal sections at different levels. Strong signal was detected in theseptum and in scattered neurons in the pontine nuclei and motor root ofthe trigeminal nerve; moderate signal was seen in the molecular layersof the hippocampus and outer layers of the retrosplenial cortex.

[2729] (5) DNA39979-1213 (PRO296)

[2730] Widespread expression in fetal in adult tissues. Expressed in avariety of fetal and adult epithelia, skeletal and cardiac muscle,developing (including retina) and adult CNS, thymic epithelium,placental villi, hepatocytes in cirrhotic and acetaminophen inducedtoxicity. Highly expressed in hypertrophic chondrocytes in developingskeletal system. The overall expression pattern, while not completelyoverlapping (not expressed in glomeruli, more widely expressed in CNS),is not disimilar to VEGF. A possible role in angiogenesis shouldtherefore be considered. Human fetal tissues examined (E12-E16 weeks)include: placenta, umbilical cord, liver, kidney, adrenals, thyroid,lungs, great vessels, stomach, small intestine, spleen, thymus,pancreas, brain, eye, spinal cord, body wall, pelvis, testis and lowerlimb. Adult human tissues examined: kidney (normal and end- stage),adrenal, spleen, lymph node, pancreas, lung, eye (inc. retina), bladder,liver (normal, cirrhotic, acute failure). Non-human primate tissuesexamined: Chimp tissues: adrenal. Rhesus Monkey tissues: cerebralcortex, hippocampus, cerebellum.

[2731] (6) DNA52594-1270 (PRO868)

[2732] Expression over neuronal cells in fetal dorsal root ganglia,spinal cord, developing enteric neurons, cortical neurons. Low levelexpression also seen in placental trophoblast. In adult tissues the onlysite where notable expression was observed was the normal adultprostate; as such it may represent a possible prostate cell surfacereceptor target antigen. Studies to further characterize the expressionin adult tissues seem warranted. Low level expression also observed inaliposarcoma. Fetal tissues examined (E12-E16 weeks) include: placenta,umbilical cord, liver, kidney, adrenals, thyroid, lungs, heart, greatvessels, oesophagus, stomach, small intestine, spleen, thymus, pancreas,brain, eye, spinal cord, body wall, pelvis and lower limb. Adult humantissues examined: liver, kidney, adrenal, myocardium, aorta, spleen,lung, skin, chondrosarcoma, eye, stomach, gastric carcinoma, colon,colonic carcinoma, renal cell carcinoma, prostate, bladder mucosa andgall bladder. Acetominophen induced liver injury and hepatic cirrhosis.Rhesus tissues examined: cerebral cortex (rm), hippocampus(rm),cerebellum. Chimp tissues examined: thyroid, parathyroid, ovary, nerve,tongue, thymus, adrenal, gastric mucosa and salivary gland. WIG-1(WISP-1), WIG-2 (WISP-2) and WIG-5 (WISP-3) expression in human breastcarcinoma and normal breast tissue, Wig-2 in lung carcinoma, and Wig-5in colon carcinoma.

[2733] (7) DNA64907-1163 (PRO1330)

[2734] In human fetal tissues there was strong specific expression overartrerial, venous, capillary and sinusoidal endothelium in all tissuesexamined, except for fetal brain. In normal adult tissues expression waslow to absent, but when present appeared expression was confined to thevasculature. Highest expression in adult tissues was observed regionallyin vessels running within the white matter of rhesus brain—thesignificance of this pattern is unclear. Elevated expression observed invasculature of many inflamed and diseased tissues, including tumorvasculature. In some of these tissues it was unclear if expression wassoley confined to vascular endothelium. In the 15 lung tumors examinedno expression was seen over the malignant epithelium, however, vascularexpression was observed in many of the tumors and adjacent lung tissue.Moderate, apparently non- specific background, was seen with this probeover hyalinised collagen and sites of tissue necrosis. In the abscenceof a sense control, however, it is not possible to be absolutely certainthat all of this signal is non- specific. Some signal, also thought tobe non-specific, was seen over the chimp gastric mucosa, transitionalcell epithelium of human adult bladder and fetal retina.

[2735] (8) DNA49624-1279 (PRO545)

[2736] Expression of the ADAM family molecule, ADAM 12 (DNA49624-1279)observed in normal human lung, lung tumor, normal colon and coloncarcinoma.

[2737] (9) DNA59294-1381 (PRO1031)

[2738] The expression of this IL17 homologue was evaluated in a panelconsisting of normal adult and fetal tissues and tissues withinflammation, predominantly chronic lymphocytic inflammation. This panelis designed to specifically evaluate the expression pattern in immunemediated inflammatory disease of novel proteins that modulate Tlymphocyte function (stimulatory or inhibitory). This protein whenexpressed as an Ig-fusion protein was immunostimulatory in a dosedependent fashion in the human mixed lymphocyte reaction (MLR); itcaused a 285% and 147% increase above the baseline stimulation indexwhen utilized at two different concentrations (1.0% and 0.1% of a 560 nMstock). Summary: expression was restricted to muscle, certain types ofsmooth muscle in the adult and in skeletal and smooth muscle in thehuman fetus. Expression in adult human was in smooth muscle of tubularorgans evaluated including colon and gall bladder. There no expressionin the smooth muscle of vessels or bronchi. No adult human skeletalmuscle was evaluated. In fetal tissues there was moderate to highdiffuse expression in skeletal muscle the axial skeleton and limbs.There was weak expression in the smooth muscle of the intestinal wallbut no expression in cardiac muscle. Adult human tissues withexpression: Colon. there was low level diffuse expression in the smoothmuscle (tunica muscularis) in 5 specimens with chronic inflammatorybowel disease. Gall bladder: there was weak to low level expression inthe smooth muscle of the gall bladder. Fetal human tissues withexpression: there was moderate diffuse expression in skeletal muscle andweak tolow expression in smooth muscle; there was no expression in fetalheart or any other fetal organ including liver, spleen, CNS, kidney,gut, lung. Human tissues with no expression: lung with chronicgranulomatous inflammation and chronic bronchitis (5 patients),peripheral nerve, prostate, heart, placenta, liver (disease multiblock),brain (cerbrum and cerebellum), tonsil (reactive hyperplasia),peripheral lymph node, thymus.

[2739] (10) DNA45416-1251 (PRO362)

[2740] The expression of this novel protein was evaluated in a varietyof human and non-human primate tissues and was found to be highlyrestricted. Expression was present only in alveolar macrophages in thelung and in Kupffer cells of the hepatic sinusoids. Expression in thesecells was significantly increased when these distinct cell populationswere activated. Though these two subpopulations of tissue macrophagesare located in different organs, they have similar biological functions.Both types of these phagocytes act as biological filters to removematerial from the blood stream or airways including pathogens, senescentcells and proteins and both are capable of secreting a wide variety ofimportant proinflammatory cytokines. In inflamed lung (7 patientsamples) expression was prominent in reactive alveolar macrophage cellpopulations defined as large, pale often vacuolated cells present singlyor in aggregates within alveoli and was weak to negative in normal,non-reactive macrophages (single scattered cells of normal size).Expression in alveolar macrophages was increased during inflammationwhen these cells were both increased in numbers and size (activated).Despite the presence of histocytes in areas of interstial inflammtionand peribronchial lymphoid hyperplasia in these tissues, expression wasrestricted to alveolar macrophages. Many of the inflamed lungs also hadsome degree of suppurative inflammation; expression was not present inneutrophilic granulocytes. In liver, there was strong expression inreactive/activated Kupffer cells in livers with acute centrilobularnecrosis (acetominophen toxicity) or fairly marked periportalinflammtion. However there was weak or no expression in Kupffer cells innormal liver or in liver with only mild inflammation or mild to moderatelobular hyperplasia/hypertrophy. Thus, as in the lung, there wasincreased expression in activated/reactive cells. There was noexpression of this molecule in histiocytes/macropahges present ininflamed bowel, hyperplastic/reactive tonsil or normal lymph node. Thelack of expression in these tissues which all contained histiocyticinflammation or resident macrophage populations strongly supportsrestricted expression to the unique macrophage subset populationsdefined as alveolar macrophage and hepatic Kupffer cells. Spleen or bonemarrow was not available for evaluation. Human tissues evaluated whichhad no detectable expression included: Inflammatory bowel disease (7patient samples with moderate to severe disease), tonsil with reactivehyperplasia, peripheral lymph node, psoriatic skin (2 patient sampleswith mild to moderate disease), heart, peripheral nerve. Chimp tissuesevaluated which had no detectable expression included: tongue, stomach,thymus.

[2741] (11) DNA52196-1348 (PRO733)

[2742] Generalized low level signal in many tissues and in many celltypes. While endothelial cell expression was observed it was not aprominent feature in either fetal, normal or diseased tissues. Humantissues: moderate expression over fetal liver (mainly hepatocytes),lung, skin, adrenal and heart. Fetal spleen, small intestine, brain andeye are negative. Adult normal kidney, bladder epithelium, lung,adrenal, pancreas, skin—all negative. Expression in adult human liver(normal and diseased), renal tubules in end-stage renal disease, adiposetissue, sarcoma, colon, renal cell carcinoma, hepatocellular carcinoma,squamous cell carcinoma. Non human primate tissues: chimp salivarygland, vessels, stomach, tongue, peripheral nerve, thymus, lymph node,thyroid and parathyroid. Rhesus spinal cord negative, cortical andhippocampal neurones positive.

Example 143 Isolation of cDNA Clones Encoding a Human PRO4993

[2743] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA85042. In some cases, the DNA85042consensus sequence derives from an intermediate consensus DNA sequencewhich was extended using repeated cycles of BLAST and phrap to extendthat intermediate consensus sequence as far as possible using thesources of EST sequences discussed above. Based on the DNA85042consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO4993.

[2744] PCR primers (forward and reverse) were synthesized:

[2745] forward PCR primer 5′-AGATGTGAAGGTGCAGGTGTGCCG-3′ (SEQ ID NO:619)

[2746] reverse PCR primer 5′-GAACATCAGCGCTCCCGGTAATTCC-3′ (SEQ IDNO:620)

[2747] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA85042 sequence which had the followingnucleotide sequence

[2748] hybridization probe

[2749] 5′-CCAGCCTTTGAATGGTACAAAGGAGAGAAGAAGCTCTTCAATGGCC-3′ (SEQ IDNO:621)

[2750] RNA for construction of the cDNA libraries was isolated fromhuman fetal brain tissue.

[2751] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for a full-length PRO4993 polypeptide(designated herein as DNA94832-2659 [FIG. 229, SEQ ID NO:6111) and thederived protein sequence for that PRO4993 polypeptide.

[2752] The full length clone identified above contained a single openreading frame with an apparent translational initiation site atnucleotide positions 305-307 and a stop signal at nucleotide positions1361-1363 (FIG. 229, SEQ ID NO:611). The predicted polypeptide precursoris 352 amino acids long, has a calculated molecular weight ofapproximately 38,429 daltons and an estimated pI of approximately 6.84.Analysis of the full-length PRO4993 sequence shown in FIG. 230 (SEQ IDNO:612) evidences the presence of a variety of important polypeptidedomains as shown in FIG. 230, wherein the locations given for thoseimportant polypeptide domains are approximate as described above. CloneDNA94832-2659 has been deposited with ATCC on Jun. 15, 1999 and isassigned ATCC deposit no. 240-PTA.

[2753] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using the ALIGN-2 sequence alignment analysis of the full-lengthsequence shown in FIG. 230 (SEQ ID NO:612), evidenced sequence identitybetween the PRO4993 amino acid sequence and the following Dayhoffsequences: P_W05152; LAMP_HUMAN; P_W05157; P_W05155; I56551; OPCM_RAT;AMAL_DROME; DMU78177_(—)1; I_(37246;) and NCA1_HUMAN.

Example 144 Isolation of cDNA Clones Encoding Human PRO1559, PRO725 andPRO739

[2754] A consensus sequence was obtained relative to a variety of ESTsequences as described in Example 1 above. Based upon an observedhomology between this consensus sequence and an EST sequence containedwithin Incyte EST clone No. 4242090, Incyte EST clone No. 4242090 waspurchased and its insert was obtained and sequenced. It was discoveredthat the insert sequence encoded a full-length protein designated hereinas PRO1559 (FIG. 232; SEQ ID NO:614). The DNA sequence of the insert(DNA68886) is shown in FIG. 231 (SEQ ID NO:613).

[2755] A cDNA sequence isolated in the amylase screen described inExample 2 above is herein designated DNA43301. The DNA43301 sequence wasthen compared to a variety of expressed sequence tag (EST) databaseswhich included public EST databases (e.g., GenBank) and a proprietaryEST DNA database (LIFESEQ™, Incyte Pharmaceuticals, Palo Alto, Calif.)to identify existing homologies. The homology search was performed usingthe computer program BLAST or BLAST2 (Altshul et al., Methods inEnzymology 266:460-480 (1996)). Those comparisons resulting in a BLASTscore of 70 (or in some cases 90) or greater that did not encode knownproteins were clustered and assembled into consensus DNA sequences withthe program “phrap” Phil Green, University of Washington, Seattle,Wash.). The consensus sequence obtained therefrom is herein designatedDNA45458. Based on the DNA45458 consensus sequence, oligonucleotideprobes were generated and used to screen a human fetal brain (LIB153)library prepared as described in paragraph 1 of Example 2 above. Thecloning vector was pRK5B (pRK5B is a precursor of pRK5D that does notcontain the SfiI site; see, Holmes et al., Science, 253:1278-1280(1991)), and the cDNA size cut was less than 2800 bp.

[2756] PCR primers (forward and reverse) were synthesized: forward PCRprimer (45458.f1) 5′-CCAAACTCACCCAGTGAGTGTGAGC-3′ (SEQ ID NO:619)reverse PCR primer (45458.r1) 5′-TGGGAAATCAGGAATGGTGTTCTCC-3′ (SEQ IDNO:620)

[2757] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA45458 sequence which had the followingnucleotide sequence

[2758] hybridization probe (45458.p1)

[2759] 5′-CTTGTTTTCACCATTGGGCTAACTTTGCTGCTAGGAGTTCAAGCCATGCC-3′ (SEQ IDNO:621)

[2760] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO725 gene usingthe probe oligonucleotide and one of the PCR primers.

[2761] A full length clone was identified that contained a single openreading frame with an apparent translational initiation site atnucleotide positions 161-163 and ending at the stop codon found atnucleotide positions 455-457 (FIG. 233; SEQ ID NO:615). The predictedpolypeptide precursor is 98 amino acids long, has a calculated molecularweight of approximately 11,081 daltons and an estimated pI ofapproximately 6.68. Analysis of the full-length PRO725 sequence shown inFIG. 234 (SEQ ID NO:616) evidences the presence of the following: asignal peptide from about amino acid 1 to about amino acid 20, apotential N-glycosylation site from about amino acid 72 to about aminoacid 75 and a tyrosine kinase phosphorylation site from about amino acid63 to about amino acid 70. Clone DNA52758-1399 has been deposited withATCC on Apr. 14, 1998 and is assigned ATCC deposit no. 209773.

[2762] Analysis of the amino acid sequence of the full-length PRO725polypeptide suggests that it possesses no significant sequencesimilarity to any known protein. However, an analysis of the Dayhoffdatabase (version 35.45 SwissProt 35) evidenced some degree of homologybetween the PRO725 amino acid sequence and the following Dayhoffsequences, POL_BLVAU, PSSP_RAT, CELC36C5_(—)7, AF019234_(—)1, I_(48862,)P_R12498, P_P10125, P_R26861, A64527 and P_W20495.

[2763] DNA52756, as shown in FIG. 235 (SEQ ID NO:617) and which encodesnative PRO739 polypeptide (FIG. 236; SEQ ID NO:618) was obtained fromGenBank.

Example 145 Identification of Receptor/Ligand Interactions

[2764] In this assay, various PRO polypeptides are tested for ability tobind to a panel of potential receptor molecules for the purpose ofidentifying receptor/ligand interactions. The identification of a ligandfor a known receptor, a receptor for a known ligand or a novelreceptor/ligand pair is useful for a variety of indications including,for example, targeting bioactive molecules (linked to the ligand orreceptor) to a cell known to express the receptor or ligand, use of thereceptor or ligand as a reagent to detect the presence of the ligand orreceptor in a composition suspected of containing the same, wherein thecomposition may comprise cells suspected of expressing the ligand orreceptor, modulating the growth of or another biological orimmunological activity of a cell known to express or respond to thereceptor or ligand, modulating the immune response of cells or towardcells that express the receptor or ligand, allowing the preparation ofagonists, antagonists and/or antibodies directed against the receptor orligand which will modulate the growth of or a biological orimmunological activity of a cell expressing the receptor or ligand, andvarious other indications which will be readily apparent to theordinarily skilled artisan.

[2765] The assay is performed as follows. A PRO polypeptide of thepresent invention suspected of being a ligand for a receptor isexpressed as a fusion protein comaning the Fc domain of human IgG (animmunoadhesin). Receptor-ligand binding is detected by allowinginteraction of the immunoadhesin polypeptide with cells (e.g. Cos cells)expressing candidate PRO polypeptide receptors and visualization ofbound immunoadhesin with fluorescent reagents directed toward the Fcfusion domain and examination by microscope. Cells expressing candidatereceptors are produced by transient transfection, in parallel, ofdefined subsets of a library of cDNA expression vectors encoding PROpolypeptides that may function as receptor molecules. Cells are thenincubated for 1 hour in the presence of the PRO polypeptideimmunoadhesin being tested for possible receptor binding. The cells arethen washed and fixed with paraformaldehyde. The cells are thenincubated with fluorescent conjugated antibody directed against the Fcportion of the PRO polypeptide immunoadhesin (e.g. FITC conjugated goatanti-human-Fc antibody). The cells are then washed again and examined bymicroscope. A positive interaction is judged by the presence offluorescent labeling of cells transfected with cDNA encoding aparticular PRO polypeptide receptor or pool of receptors and an absenceof similar fluorescent labeling of similarly prepared cells that havebeen transfected with other cDNA or pools of cDNA. If a defined pool ofcDNA expression vectors is judged to be positive for interaction with aPRO polypeptide immunoadhesin, the individual cDNA species that comprisethe pool are tested individually (the pool is “broken down”) todetermine the specific cDNA that encodes a receptor able to interactwith the PRO polypeptide immunoadhesin.

[2766] In another embodiment of this assay, an epitope-tagged potentialligand PRO polypeptide (e.g. 8 histidine “His” tag) is allowed tointeract with a panel of potential receptor PRO polypeptide moleculesthat have been expressed as fusions with the Fc domain of human IgG(immunoadhesins). Following a 1 hour co-incubation with the epitopetagged PRO polypeptide, the candidate receptors are eachimmunoprecipitated with protein A beads and the beads are washed.Potential ligand interaction is determined by western blot analysis ofthe immunoprecipitated complexes with antibody directed towards theepitope tag. An interaction is judged to occur if a band of theanticipated molecular weight of the epitope tagged protein is observedin the western blot analysis with a candidate receptor, but is notobserved to occur with the other members of the panel of potentialreceptors.

[2767] Using these assays, the following receptor/ligand interactionshave been herein identified: PRO337 binds to PRO4993, PRO1559 binds toPRO725, PRO1559 binds to PRO700 and PRO1559 binds to PRO739.

[2768] Deposit of Material

[2769] The following materials have been deposited with the AmericanType Culture Collection, 12301 Parklawn Drive, Rockville, Md., USA(ATCC): Material ATCC Dep. No. Deposit Date DNA39987-1184 ATCC 209786April 21, 1998 DNA40625-1189 ATCC 209788 April 21, 1998 DNA23318-1211ATCC 209787 April 21, 1998 DNA39979-1213 ATCC 209789 April 21, 1998DNA40594-1233 ATCC 209617 February 5, 1998 DNA45416-1251 ATCC 209620February 5, 1998 DNA45419-1252 ATCC 209616 February 5, 1998DNA52594-1270 ATCC 209679 March 17, 1998 DNA45234-1277 ATCC 209654 March5, 1998 DNA49624-1279 ATCC 209655 March 5, 1998 DNA48309-1280 ATCC209656 March 5, 1998 DNA46776-1284 ATCC 209721 March 31, 1998DNA50980-1286 ATCC 209717 March 31, 1998 DNA50913-1287 ATCC 209716 March31, 1998 DNA50914-1289 ATCC 209722 March 31, 1998 DNA48296-1292 ATCC209668 March 11, 1998 DNA32284-1307 ATCC 209670 March 11, 1998DNA36343-1310 ATCC 209718 March 31, 1998 DNA40571-1315 ATCC 209784 April21, 1998 DNA41386-1316 ATCC 209703 March 26, 1998 DNA44194-1317 ATCC209808 April 28, 1998 DNA45415-1318 ATCC 209810 April 28, 1998DNA44189-1322 ATCC 209699 March 26, 1998 DNA48304-1323 ATCC 209811 April28, 1998 DNA49152-1324 ATCC 209813 April 28, 1998 DNA49646-1327 ATCC209705 March 26, 1998 DNA49631-1328 ATCC 209806 April 28, 1998DNA49645-1347 ATCC 209809 April 28, 1998 DNA45493-1349 ATCC 209805 April28, 1998 DNA48227-1350 ATCC 209812 April 28, 1998 DNA41404-1352 ATCC209844 May 6, 1998 DNA44196-1353 ATCC 209847 May 6, 1998 DNA52187-1354ATCC 209845 May 6, 1998 DNA48328-1355 ATCC 209843 May 6, 1998DNA56352-1358 ATCC 209846 May 6, 1998 DNA53971-1359 ATCC 209750 April 7,1998 DNA50919-1361 ATCC 209848 May 6, 1998 DNA44179-1362 ATCC 209851 May6, 1998 DNA54002-1367 ATCC 209754 April 7, 1998 DNA53906-1368 ATCC209747 April 7, 1998 DNA52185-1370 ATCC 209861 May 14, 1998DNA53977-1371 ATCC 209862 May 14, 1998 DNA57253-1382 ATCC 209867 May 14,1998 DNA58847-1383 ATCC 209879 May 20, 1998 DNA58747-1384 ATCC 209868May 14, 1998 DNA57689-1385 ATCC 209869 May 14, 1998 DNA23330-1390 ATCC209775 April 14, 1998 DNA26847-1395 ATCC 209772 April 14, 1998DNA53974-1401 ATCC 209774 April 14, 1998 DNA57039-1402 ATCC 209777 April14, 1998 DNA57033-1403 ATCC 209905 May 27, 1998 DNA34353-1428 ATCC209855 May 12, 1998 DNA45417-1432 ATCC 209910 May 27, 1998 DNA39523-1192ATCC 209424 October 31, 1997 DNA44205-1285 ATCC 209720 March 31, 1998DNA5O911-1288 ATCC 209714 March 31, 1998 DNA48329-1290 ATCC 209785 April21, 1998 DNA48306-1291 ATCC 209911 May 27, 1998 DNA48336-1309 ATCC209669 March 11, 1998 DNA44184-1319 ATCC 209704 March 26, 1998DNA48314-1320 ATCC 209702 March 26, 1998 DNA48333-1321 ATCC 209701 March26, 1998 DNA50920-1325 ATCC 209700 March 26, 1998 DNA50988-1326 ATCC209814 April 28, 1998 DNA48331-1329 ATCC 209715 March 31, 1998DNA30867-1335 ATCC 209807 April 28, 1998 DNA55737-1345 ATCC 209753 April7, 1998 DNA49829-1346 ATCC 209749 April 7, 1998 DNA52196-1348 ATCC209748 April 7, 1998 DNA56965-1356 ATCC 209842 May 6, 1998 DNA56405-1357ATCC 209849 May 6, 1998 DNA57530-1375 ATCC 209880 May 20, 1998DNA56439-1376 ATCC 209864 May 14, 1998 DNA56409-1377 ATCC 209882 May 20,1998 DNAS6112-1379 ATCC 209883 May 20, 1998 DNA56045-1380 ATCC 209865May 14, 1998 DNA59294-1381 ATCC 209866 May 14, 1998 DNA56433-1406 ATCC209857 May 12, 1998 DNA53912-1457 ATCC 209870 May 14, 1998 DNA50921-1458ATCC 209859 May 12, 1998 DNA29101-1122 ATCC 209653 March 5, 1998DNA40021-1154 ATCC 209389 October 17, 1997 DNA42663-1154 ATCC 209386October 17, 1997 DNA30943-1-1163-1 ATCC 209791 April 21, 1998DNA64907-1163-1 ATCC 203242 September 9, 1998 DNA64908-1163-1 ATCC203243 September 9, 1998 DNA39975-1210 ATCC 209783 April 21, 1998DNA43316-1237 ATCC 209487 November 21, 1997 DNA55800-1263 ATCC 209680March 17, 1998 DNA94832-2659 240-PTA June 15, 1999 DNA52758-1399 ATCC209773 April 14, 1998

[2770] These deposit were made under the provisions of the BudapestTreaty on the International Recognition of the Deposit of Microorganismsfor the Purpose of Patent Procedure and the Regulations thereunder(Budapest Treaty). This assures maintenance of a viable culture of thedeposit for 30 years from the date of deposit. The deposits will be madeavailable by ATCC under the terms of the Budapest Treaty, and subject toan agreement between Genentech, Inc. and ATCC, which assures permanentand unrestricted availability of the progeny of the culture of thedeposit to the public upon issuance of the pertinent U.S. patent or uponlaying open to the public of any U.S. or foreign patent application,whichever comes first, and assures availability of the progeny to onedetermined by the U.S. Commissioner of Patents and Trademarks to beentitled thereto according to 35 USC § 122 and the Commissioner's rulespursuant thereto (including 37 CFR § 1.14 with particular reference to886 OG 638).

[2771] The assignee of the present application has agreed that if aculture of the materials on deposit should die or be lost or destroyedwhen cultivated under suitable conditions, the materials will bepromptly replaced on notification with another of the same. Availabilityof the deposited material is not to be construed as a license topractice the invention in contravention of the rights granted under theauthority of any government in accordance with its patent laws.

[2772] The foregoing written specification is considered to besufficient to enable one skilled in the art to practice the invention.The present invention is not to be limited in scope by the constructdeposited, since the deposited embodiment is intended as a singleillustration of certain aspects of the invention and any constructs thatare functionally equivalent are within the scope of this invention. Thedeposit of material herein does not constitute an admission that thewritten description herein contained is inadequate to enable thepractice of any aspect of the invention, including the best modethereof, nor is it to be construed as limiting the scope of the claimsto the specific illustrations that it represents. Indeed, variousmodifications of the invention in addition to those shown and describedherein will become apparent to those skilled in the art from theforegoing description and fall within the scope of the appended claims.

APPENDIX A

[2773] I hereby claim the benefit under Title 35, United States Code,§119(e) of any United States provisional applications listed below:

[2774] 1. 60/062250 filed Oct. 17, 1997

[2775] 2. 60/064249 filed Nov. 3, 1997

[2776] 3. 60/065311 filed Nov. 13, 1997

[2777] 4. 60/066364 filed Nov. 21, 1997

[2778] 5. 60/077450 filed Mar. 10, 1998

[2779] 6. 60/077632 filed Mar. 11, 1998

[2780] 7. 60/077641 filed Mar. 11, 1998

[2781] 8. 60/077649 filed Mar. 11, 1998

[2782] 9. 60/077791 filed Mar. 12, 1998

[2783] 10. 60/078004 filed Mar. 13, 1998

[2784] 11. 60/078886 filed Mar. 20, 1998

[2785] 12. 60/078936 filed Mar. 20, 1998

[2786] 13. 60/078910 filed Mar. 20, 1998

[2787] 14. 60/078939 filed Mar. 20, 1998

[2788] 15. 60/079294 filed Mar. 25, 1998

[2789] 16. 60/079656 filed Mar. 26, 1998

[2790] 17. 60/079664 filed Mar. 27, 1998

[2791] 18. 60/079689 filed Mar. 27, 1998

[2792] 19. 60/079663 filed Mar. 27, 1998

[2793] 20. 60/079728 filed Mar. 27, 1998

[2794] 21. 60/079786 filed Mar. 27, 1998

[2795] 22. 60/079920 filed Mar. 30, 1998

[2796] 23. 60/079923 filed Mar. 30, 1998

[2797] 24. 60/080105 filed Mar. 31, 1998

[2798] 25. 60/080107 filed Mar. 31, 1998

[2799] 26. 60/080165 filed Mar. 31, 1998

[2800] 27. 60/080194 filed Mar. 31, 1998

[2801] 28. 60/080327 filed Apr. 1, 1998

[2802] 29. 60/080328 filed Apr. 1, 1998

[2803] 30. 60/080333 filed Apr. 1, 1998

[2804] 31. 60/080334 filed Apr. 1, 1998

[2805] 32. 60/081070 filed Apr. 8, 1998

[2806] 33. 60/081049 filed Apr. 8, 1998

[2807] 34. 60/081071 filed Apr. 8, 1998

[2808] 35. 60/081195 filed Apr. 9, 1998

[2809] 36. 60/081203 filed Apr. 9, 1998

[2810] 37. 60/081229 filed Apr. 9, 1998

[2811] 38. 60/081955 filed Apr. 15, 1998

[2812] 39. 60/081817 filed Apr. 15, 1998

[2813] 40. 60/081819 filed Apr. 15, 1998

[2814] 41. 60/081952 filed Apr. 15, 1998

[2815] 42. 60/081838 filed Apr. 15, 1998

[2816] 43. 60/082568 filed Apr. 21, 1998

[2817] 44. 60/082569 filed Apr. 21, 1998

[2818] 45. 60/082704 filed Apr. 22, 1998

[2819] 46. 60/082804 filed Apr. 22, 1998

[2820] 47. 60/082700 filed Apr. 22, 1998

[2821] 48. 60/082797 filed Apr. 22, 1998

[2822] 49. 60/082796 filed Apr. 23, 1998

[2823] 50. 60/083336 filed Apr. 27, 1998

[2824] 51. 60/083322 filed Apr. 28, 1998

[2825] 52. 60/083392 filed Apr. 29, 1998

[2826] 53. 60/083495 filed Apr. 29, 1998

[2827] 54. 60/083496 filed Apr. 29, 1998

[2828] 55. 60/083499 filed Apr. 29, 1998

[2829] 56. 60/083545 filed Apr. 29, 1998

[2830] 57. 60/083554 filed Apr. 29, 1998

[2831] 58. 60/083558 filed Apr. 29, 1998

[2832] 59. 60/083559 filed Apr. 29, 1998

[2833] 60. 60/083500 filed Apr. 29, 1998

[2834] 61. 60/083742 filed Apr. 30, 1998

[2835] 62. 60/084366 filed May 5, 1998

[2836] 63. 60/084414 filed May 6, 1998

[2837] 64. 60/084441 filed May 6, 1998

[2838] 65. 60/084637 filed May 7, 1998

[2839] 66. 60/084639 filed May 7, 1998

[2840] 67. 60/084640 filed May 7, 1998

[2841] 68. 60/084598 filed May 7, 1998

[2842] 69. 60/084600 filed May 7, 1998

[2843] 70. 60/084627 filed May 7, 1998

[2844] 71. 60/084643 filed May 7, 1998

[2845] 72. 60/085339 filed May 13, 1998

[2846] 73. 60/085338 filed May 13, 1998

[2847] 74. 60/085323 filed May 13, 1998

[2848] 75. 60/085582 filed May 15, 1998

[2849] 76. 60/085700 filed May 15, 1998

[2850] 77. 60/085689 filed May 15, 1998

[2851] 78. 60/085579 filed May 15, 1998

[2852] 79. 60/085580 filed May 15, 1998

[2853] 80. 60/085573 filed May 15, 1998

[2854] 81. 60/085704 filed May 15, 1998

[2855] 82. 60/085697 filed May 15, 1998

[2856] 83. 60/086023 filed May 18, 1998

[2857] 84. 60/086430 filed May 22, 1998

[2858] 85. 60/086392 filed May 22, 1998

[2859] 86. 60/086486 filed May 22, 1998

[2860] 87. 60/086414 filed May 22, 1998

[2861] 88. 60/087208 filed May 28, 1998

[2862] 89. 60/087106 filed May 28, 1998

[2863] 90. 60/087098 filed May 28, 1998

[2864] 91. 60/091010 filed Jun. 26, 1998

[2865] 92. 60/690863 filed Jun. 26, 1998

[2866] 93. 60/091359 filed Jul. 1, 1998

[2867] 94. 60/094651 filed Jul. 30, 1998

[2868] 95. 60/100038 filed Sep. 11, 1998

[2869] 96. 60/109304 filed Nov. 20, 1998

[2870] 97. 60/113296 filed Dec. 22, 1998

[2871] 98. 60/113621 filed Dec. 23, 1998

[2872] 99. 60/123957 filed Mar. 12, 1999

[2873] 100. 60/126773 filed Mar. 29, 1999

[2874] 101. 60/130232 filed Apr. 21, 1999

[2875] 102. 60/131022 filed Apr. 26, 1999

[2876] 103. 60/131445 filed Apr. 28, 1999

[2877] 104. 60/134287 filed May 14, 1999

[2878] 105. 60/139557 filed Jun. 16, 1999

[2879] 106. 60/141037 filed Jun. 23, 1999

[2880] 107. 60/142680 filed Jul. 7, 1999

[2881] 108. 60/145698 filed Jul. 26, 1999

[2882] 109. 60/146222 filed Jul. 28, 1999

[2883] 110. 60/162506 filed Oct. 29, 1999

APPENDIX B

[2884] I hereby claim the benefit under Title 35, United States Code,§120 of any United States applications listed below:

[2885] U.S. patent applications

[2886] 1. 09/040220 filed Mar. 17, 1998

[2887] 2. 09/105413 filed Jun. 26, 1998

[2888] 3. 09/168978 filed Oct. 7, 1998

[2889] 4. 09/184216 filed Nov. 2,1998

[2890] 5. 09/187368 filed Nov. 6, 1998

[2891] 6. 09/202054 filed Dec. 7, 1998

[2892] 7. 09/218517 filed Dec. 22,1998

[2893] 8. 09/254465 filed Mar. 5, 1999

[2894] 9. 09/265686 filed Mar. 10, 1999

[2895] 10. 09/267213 filed Mar. 12, 1999

[2896] 11. 09/284291 filed Apr. 12,1999

[2897] 12. 09/311832 filed May 14, 1999

[2898] 13. 09/380137 filed Aug. 25, 1999

[2899] 14. 09/380138 filed Aug. 25, 1999

[2900] 15. 09/380142 filed Aug. 25, 1999

[2901] 16. 09/709238 filed Nov. 8, 2000

[2902] 17. 09/723749 filed Nov. 27, 2000

[2903] 18. 09/747259 filed Dec. 20, 2000

[2904] 19. 09/816744 filed Mar. 22,2001

[2905] 20. 09/816920 filed Mar. 22, 2001

[2906] 21. 09/854280 filed May 10, 2001

[2907] 22. 09/854208 filed May 10, 2001

[2908] 23. 09/872035 filed Jun. 1, 2001

[2909] 24. 09/874503 filed Jun. 5, 2001

[2910] 25. 09/882636 filed Jun. 14, 2001

[2911] 26. 09/886342 filed Jun. 19, 2001

[2912] PCT APPLICATIONS

[2913] 1. PCT/US98/21141 filed Oct. 7, 1998

[2914] 2. PCT/US98/24855 filed Nov. 20, 1998

[2915] 3. PCT/US99/00106 filed Jan. 5, 1999

[2916] 4. PCT/US99/05028 filed Mar. 8, 1999

[2917] 5. PCT/US99/05190 filed Mar. 10, 1999

[2918] 6. PCT/US99/10733 filed May 14,1999

[2919] 7. PCT/US99/12252 filed Jun. 2, 1999

[2920] 8. PCT/US99/28313 filed Nov. 30, 1999

[2921] 9. PCT/US99/28551 filed Dec. 2, 1999

[2922] 10. PCT/US99/28565 filed Dec. 2, 1999

[2923] 11. PCT/US99/30095 filed Dec. 16, 1999

[2924] 12. PCT/US99/31243 filed Dec. 30, 1999

[2925] 13. PCT/US99/31274 filed Dec. 30, 1999

[2926] 14. PCT/US00/00219 filed Jan. 5, 2000

[2927] 15. PCT/US00/00277 filed Jan. 6, 2000

[2928] 16. PCT/US00/00376 filed Jan. 6, 2000

[2929] 17. PCT/US00/03565 filed Feb. 11, 2000

[2930] 18. PCT/US00/04341 filed Feb. 18, 2000

[2931] 19. PCT/US00/05841 filed Mar. 2, 2000

[2932] 20. PCT/US00/07532 filed Mar. 21, 2000

[2933] 21. PCT/US00/05004 filed Feb. 24, 2000

[2934] 22. PCT/US00/06319 filed Mar. 10, 2000

[2935] 23. PCT/US00/08439 filed Mar. 30,2000

[2936] 24. PCT/US00/13705 filed May 17, 2000

[2937] 25. PCT/US00/14042 filed May 22,2000

[2938] 26. PCT/US00/14941 filed May 30, 2000

[2939] 27. PCT/US00/15264 filed Jun. 2,2000

[2940] 28. PCT/US00/20710 filed Jul. 28, 2000

[2941] 29. PCT/US00/23328 filed Aug. 24, 2000

[2942] 30. PCT/US00/32678 filed Dec. 1, 2000

[2943] 31. PCT/US00/34956 filed Dec. 20, 2000

[2944] 32. PCT/US01/06520 filed Feb. 28, 2001

[2945] 33. PCT/US01/09552 filed Mar. 22, 2001

[2946] 34. PCT/US01/17092 filed May 25, 2001

[2947] 35. PCT/US01/17800 filed Jun. 1, 2001

[2948] 36. PCT/US01/19692 filed Jun. 20,2001

[2949] 37. PCT/US01/21066 filed Jun. 29, 2001

[2950] 38. PCT/US01/21735 filed Jul. 9, 2001

0 SEQUENCE LISTING The patent application contains a lengthy “SequenceListing” section. A copy of the “Sequence Listing” is available inelectronic form from the USPTO web site(http://seqdata.uspto.gov/sequence.html?DocID=20030199435). Anelectronic copy of the “Sequence Listing” will also be available fromthe USPTO upon request and payment of the fee set forth in 37 CFR1.19(b)(3).

What is claimed is:
 1. Isolated nucleic acid having at least 80% nucleicacid sequence identity to a nucleotide sequence that encodes an aminoacid sequence selected from the group consisting of the amino acidsequence shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:7), FIG. 9(SEQ ID NO:19), FIG. 11 (SEQ ID NO:28), FIG. 15 (SEQ ID NO:36), FIG. 20(SEQ ID NO:45), FIG. 22 (SEQ ID NO:52), FIG. 24 (SEQ ID NO:59), FIG. 26(SEQ ID NO:64), FIG. 28 (SEQ ID NO:69), FIG. 30 (SEQ ID NO:74), FIG. 33(SEQ ID NO:85), FIG. 35 (SEQ ID NO:90), FIG. 37 (SEQ ID NO:97), FIG. 39(SEQ ID NO:102), FIG. 41 (SEQ ID NO:109), FIG. 43 (SEQ ID NO:114), FIG.45 (SEQ ID NO:119), FIG. 47 (SEQ ID NO:124), FIG. 49 (SEQ ID NO:132),FIG. 51 (SEQ ID NO:137), FIG. 53 (SEQ ID NO:145), FIG. 55 (SEQ IDNO:150), FIG. 59 (SEQ ID NO:157), FIG. 61 (SEQ ID NO:162j, FIG. 63 (SEQID NO:169), FIG. 66 (SEQ ID NO:178), FIG. 68 (SEQ ID NO:183), FIG. 70(SEQ ID NO:190), FIG. 73 (SEQ ID NO:196), FIG. 75 (SEQ ID NO:206), FIG.77 (SEQ ID NO:211), FIG. 79 (SEQ ID NO:216), FIG. 81 (SEQ ID NO:221),FIG. 83 (SEQ ID NO:226), FIG. 85 (SEQ ID NO:231), FIG. 87 (SEQ IDNO:236), FIG. 89 (SEQ ID NO:245), FIG. 91 (SEQ ID NO:254), FIG. 93 (SEQID NO:259), FIG. 95 (SEQ ID NO:264), FIG. 98 (SEQ ID NO:270), FIG. 109(SEQ ID NO:284), FIG. 118 (SEQ ID NO:296), FIG. 120 (SEQ ID NO:301),FIG. 122 (SEQ ID NO:303), FIG. 125 (SEQ ID NO:309), FIG. 129 (SEQ IDNO:322), FIG. 132 (SEQ ID NO:330), FIG. 136 (SEQ ID NO:337), FIG. 139(SEQ ID NO:346), FIG. 142 (SEQ ID NO:352), FIG. 145 (SEQ ID NO:358),FIG. 147 (SEQ ID NO:363), FIG. 149 (SEQ ID NO:370), FIG. 151 (SEQ IDNO:375), FIG. 153 (SEQ ID NO:380), FIG. 155 (SEQ ID NO:385), FIG. 157(SEQ ID NO:390), FIG. 159 (SEQ ID NO:395), FIG. 161 (SEQ ID NO-.400),FIG. 163 (SEQ ID NO:405), FIG. 165 (SEQ ID NO:410), FIG. 167 (SEQ IDNO:415), FIG. 169 (SEQ ID NO:420), FIG. 171 (SEQ ID NO:425), FIG. 173(SEQ ID NO:430), FIG. 177 (SEQ ID NO:437), FIG. 179 (SEQ ID NO:442),FIG. 181 (SEQ ID NO:447), FIG. 183 (SEQ ID NO:452), FIG. 185 (SEQ IDNO:454), FIG. 187 (SEQ ID NO:456), FIG. 190 (SEQ ID NO:459), FIG. 192(SEQ ID NO:464), FIG. 194 (SEQ ID NO:466), FIG. 196 (SEQ ID NO:468),FIG. 198 (SEQ ID NO:470), FIG. 200 (SEQ ID NO:472), FIG. 202 (SEQ IDNO:477), FIG. 204 (SEQ ID NO:483), FIG. 207 (SEQ ID NO:488), FIG. 209(SEQ ID NO:496), FIG. 211 (SEQ ID NO:498), FIG. 213 (SEQ ID NO:506),FIG. 215 (SEQ ID NO:508), FIG. 217 (SEQ ID NO:510), FIG. 219 (SEQ IDNO:515), FIG. 222 (SEQ ID NO:523), FIG. 225 (SEQ ID NO:526), FIG. 230(SEQ ID NO:612), FIG. 232 (SEQ ID NO:614), FIG. 234 (SEQ ID NO:616) andFIG. 236 (SEQ ID NO:618).
 2. Isolated nucleic acid having at least 80%nucleic acid sequence identity to a nucleotide sequence selected fromthe group consisting of the nucleotide sequence shown in FIG. 1 (SEQ IDNO:1), FIG. 3 (SEQ ID NO:6), FIG. 8 (SEQ ID NO:18), FIG. 10 (SEQ IDNO:27), FIG. 14 (SEQ ID NO:35), FIG. 19 (SEQ ID NO:44), FIG. 21 (SEQ IDNO:51), FIG. 23 (SEQ ID NO:58), FIG. 25 (SEQ ID NO:63), FIG. 27 (SEQ IDNO:68), FIG. 29 (SEQ ID NO:73), FIG. 32 (SEQ ID NO:84), FIG. 34 (SEQ IDNO:89), FIG. 36 (SEQ ID NO:96), FIG. 38 (SEQ ID NO:101), FIG. 40 (SEQ IDNO:108), FIG. 42 (SEQ ID NO:113), FIG. 44 (SEQ ID NO:118), FIG. 46 (SEQID NO:123), FIG. 48 (SEQ ID NO:131), FIG. 50 (SEQ ID NO:136), FIG. 52(SEQ ID NO:144), FIG. 54 (SEQ ID NO:149), FIG. 58 (SEQ ID NO:156), FIG.60 (SEQ ID NO:161), FIG. 62 (SEQ ID NO:168), FIG. 65 (SEQ ID NO:177),FIG. 67 (SEQ ID NO:182), FIG. 69 (SEQ ID NO:189), FIG. 72 (SEQ IDNO:195), FIG. 74 (SEQ ID NO:205), FIG. 76 (SEQ ID NO:210), FIG. 78 (SEQID NO:215), FIG. 80 (SEQ ID NO:220), FIG. 82 (SEQ ID NO:225), FIG. 84(SEQ ID NO:230), FIG. 86 (SEQ ID NO:235), FIG. 88 (SEQ ID NO:244), FIG.90 (SEQ ID NO:253), FIG. 92 (SEQ ID NO:258), FIG. 94 (SEQ ID NO:263),FIG. 97 (SEQ ID NO:269), FIG. 108 (SEQ ID NO:283), FIG. 117 (SEQ IDNO:295), FIG. 119 (SEQ ID NO:300), FIG. 121 (SEQ ID NO:302), FIG. 124(SEQ ID NO:308), FIG. 128 (SEQ ID NO:321), FIG. 131 (SEQ ID NO:329),FIG. 135 (SEQ ID NO:336), FIG. 138 (SEQ ID NO:345), FIG. 141 (SEQ IDNO:351), FIG. 144 (SEQ ID NO:357), FIG. 146 (SEQ ID NO:362), FIG. 148(SEQ ID NO:369), FIG. 150 (SEQ ID NO:374), FIG. 152 (SEQ ID NO:379),FIG. 154 (SEQ ID NO:384), FIG. 156 (SEQ ID NO:389), FIG. 158 (SEQ IDNO:394), FIG. 160 (SEQ ID NO:399), FIG. 162 (SEQ ID NO:404), FIG. 164(SEQ ID NO:409), FIG. 166 (SEQ ID NO:414), FIG. 168 (SEQ ID NO:419),FIG. 170 (SEQ ID NO:424), FIG. 172 (SEQ ID NO:429), FIG. 176 (SEQ IDNO:436), FIG. 178 (SEQ ID NO:441), FIG. 180 (SEQ ID NO:446), FIG. 182(SEQ ID NO:451), FIG. 184 (SEQ ID NO:453), FIG. 186 (SEQ ED NO:455),FIG. 189 (SEQ ID NO:458), FIG. 191 (SEQ ID NO:463), FIG. 193 (SEQ IDNO:465), FIG. 195 (SEQ ID NO:467), FIG. 197 (SEQ ID NO:469), FIG. 199(SEQ ID NO:471), FIG. 201 (SEQ ID NO:476), FIG. 203 (SEQ ID NO:482),FIG. 206 (SEQ ID NO:487), FIG. 208 (SEQ ID NO:495), FIG. 210 (SEQ IDNO:497), FIG. 212 (SEQ ID) NO:505), FIG. 214 (SEQ ID NO:507), FIG. 216(SEQ ID NO:509), FIG. 218 (SEQ ID NO:514), FIG. 221 (SEQ ID NO:522),FIG. 224 (SEQ ID NO:525), FIG. 229 (SEQ ID NO:611), FIG. 231 (SEQ IDNO:613), FIG. 233 (SEQ ID NO:615) and FIG. 235 (SEQ ID NO:617). 3.Isolated nucleic acid having at least 80% nucleic acid sequence identityto a nucleotide sequence selected from the group consisting of thefull-length coding sequence of the nucleotide sequence shown in FIG. I(SEQ ID NO:1), FIG. 3 (SEQ ID NO:6), FIG. 8 (SEQ ID NO:18), FIG. 10 (SEQID NO:27), FIG. 14 (SEQ ID NO:35), FIG. 19 (SEQ ID NO:44), FIG. 21 (SEQID NO:51), FIG. 23 (SEQ ID NO:58), FIG. 25 (SEQ ID NO:63), FIG. 27 (SEQID NO:68), FIG. 29 (SEQ ID NO:73), FIG. 32 (SEQ ID NO:84), FIG. 34 (SEQID NO:89), FIG. 36 (SEQ ID NO:96), FIG. 38 (SEQ ID NO:101), FIG. 40 (SEQID NO:108), FIG. 42 (SEQ ID NO:113), FIG. 44 (SEQ ID NO:118), FIG. 46(SEQ ID NO:123), FIG. 48 (SEQ ID NO:131), FIG. 50 (SEQ ID NO:136), FIG.52 (SEQ ID NO:144), FIG. 54 (SEQ ID NO:149), FIG. 58 (SEQ ID NO:156),FIG. 60 (SEQ ID NO:161), FIG. 62 (SEQ ID NO:168), FIG. 65 (SEQ IDNO:177), FIG. 67 (SEQ ID NO:182), FIG. 69 (SEQ ID NO:189), FIG. 72 (SEQID NO:195), FIG. 74 (SEQ ID NO:205), FIG. 76 (SEQ ID NO:210), FIG. 78(SEQ ID NO:215), FIG. 80 (SEQ ID NO:220), FIG. 82 (SEQ ID NO:225), FIG.84 (SEQ ID NO:230), FIG. 86 (SEQ ID NO:235), FIG. 88 (SEQ ED NO:244),FIG. 90 (SEQ ID NO:253), FIG. 92 (SEQ ID NO:258), FIG. 94 (SEQ IDNO:263), FIG. 97 (SEQ ID NO:269), FIG. 108 (SEQ ID NO:283), FIG. 117(SEQ ID NO:295), FIG. 119 (SEQ ID NO:300), FIG. 121 (SEQ ID NO:302),FIG. 124 (SEQ ID NO:308), FIG. 128 (SEQ ID NO:321), FIG. 131 (SEQ IDNO:329), FIG. 135 (SEQ I) NO:336), FIG. 138 (SEQ H) NO:345), FIG. 141(SEQ ID NO:351), FIG. 144 (SEQ ID NO:357), FIG. 146 (SEQ ID NO:362),FIG. 148 (SEQ ID NO:369), FIG. 150 (SEQ ID NO:374), FIG. 152 (SEQ IDNO:379), FIG. 154 (SEQ ID NO:384), FIG. 156 (SEQ ID NO:389), FIG. 158(SEQ ID NO:394), FIG. 160 (SEQ ID NO:399), FIG. 162 (SEQ ID NO:404),FIG. 164 (SEQ ID NO:409), FIG. 166 (SEQ ID NO:414), FIG. 168 (SEQ IDNO:419), FIG. 170 (SEQ ID NO:424), FIG. 172 (SEQ ID NO:429), FIG. 176(SEQ ID NO:436), FIG. 178 (SEQ ID NO:441), FIG. 180 (SEQ ID NO:446),FIG. 182 (SEQ ID NO:451), FIG. 184 (SEQ ID NO:453), FIG. 186 (SEQ IDNO:455), FIG. 189 (SEQ ID NO:458), FIG. 191 (SEQ ID NO:463), FIG. 193(SEQ ID NO:465), FIG. 195 (SEQ ID NO:467), FIG. 197 (SEQ ID NO:469),FIG. 199 (SEQ ID NO:471), FIG. 201 (SEQ D NO:476), FIG. 203 (SEQ IDNO:482), FIG. 206 (SEQ ID NO:487), FIG. 208 (SEQ ID NO:495), FIG. 210(SEQ ID NO:497), FIG. 212 (SEQ ID NO:505), FIG. 214 (SEQ ID NO:507),FIG. 216 (SEQ ID NO:509), FIG. 218 (SEQ ID NO:514), FIG. 221 (SEQ IDNO:522), FIG. 224 (SEQ ID NO:525), FIG. 229 (SEQ ID NO:611), FIG. 231(SEQ ID NO:613), FIG. 233 (SEQ ID NO:615) and FIG. 235 (SEQ ID NO:617).4. Isolated nucleic acid having at least 80% nucleic acid sequenceidentity to the full-length coding sequence of the DNA deposited underATCC accession number ATCC 209791, ATCC 209786, ATCC 209788, ATCC209787, ATCC 209789, ATCC 209617, ATCC 209620, ATCC 209616, ATCC 209679,ATCC 209654, ATCC 209655, ATCC 209656, ATCC 209721, ATCC 209717, ATCC209716, ATCC 209722, ATCC 209668, ATCC 209670, ATCC 209718, ATCC 209784,ATCC 209703, ATCC 209808, ATCC 209810, ATCC 209699, ATCC 209811, ATCC209813, ATCC 209705, ATCC 209806, ATCC 209809, ATCC 209805, ATCC 209812,ATCC 209844, ATCC 209847, ATCC 209845, ATCC 209843, ATCC 209846, ATCC209750, ATCC 209848, ATCC 209851, ATCC 209754, ATCC 209747, ATCC 209861,ATCC 209862, ATCC 209867, ATCC 209879, ATCC 209868, ATCC 209869, ATCC209775, ATCC 209772, ATCC 209774, ATCC 209777, ATCC 209905, ATCC 209855,ATCC 209910, ATCC 209424, ATCC 209720, ATCC 209714, ATCC 209785, ATCC209911, ATCC 209669, ATCC 209704, ATCC 209702, ATCC 209701, ATCC 209700,ATCC 209814, ATCC 209715, ATCC 209807, ATCC 209753, ATCC 209749, ATCC209748, ATCC 209842, ATCC 209849, ATCC 209880, ATCC 209864, ATCC 209882,ATCC 209883, ATCC 209865, ATCC 209866, ATCC 209857, ATCC 209870, ATCC209859, ATCC 209653, ATCC 209389, ATCC 209386, ATCC 203242, ATCC 203243,ATCC 209783, ATCC 209487, ATCC 209680, 240-PTA or ATCC
 209773. 5. Avector comprising the nucleic acid of any one of claims 1 to
 4. 6. Thevector of claim 5 operably linked to control sequences recognized by ahost cell transformed with the vector.
 7. A host cell comprising thevector of claim
 5. 8. The host cell of claim 7, wherein said cell is aCHO cell.
 9. The host cell of claim 7, wherein said cell is an E. coli.10. The host cell of claim 7, wherein said cell is a yeast cell.
 11. Aprocess for producing a PRO polypeptides comprising culturing the hostcell of claim 7 under conditions suitable for expression of said PROpolypeptide and recovering said PRO polypeptide from the cell culture.12. An isolated polypeptide having at least 80% amino acid sequenceidentity to an amino acid sequence selected from the group consisting ofthe amino acid sequence shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ IDNO:7), FIG. 9 (SEQ ID NO:19), FIG. 11 (SEQ ID NO:28), FIG. 15 (SEQ IDNO:36), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:52), FIG. 24 (SEQ IDNO:59), FIG. 26 (SEQ ID NO:64), FIG. 28 (SEQ ID NO:69), FIG. 30 (SEQ IDNO:74), FIG. 33 (SEQ ID NO:85), FIG. 35 (SEQ ID NO:90), FIG. 37 (SEQ IDNO:97), FIG. 39 (SEQ ID NO:102), FIG. 41 (SEQ ID NO:109), FIG. 43 (SEQID NO:114), FIG. 45 (SEQ ID NO:119), FIG. 47 (SEQ ID NO:124), FIG. 49(SEQ ID NO:132), FIG. 51 (SEQ ID NO:137), FIG. 53 (SEQ ID NO:145), FIG.55 (SEQ ID NO:150), FIG. 59 (SEQ ID NO:157), FIG. 61 (SEQ ID NO:162),FIG. 63 (SEQ ID NO:169), FIG. 66 (SEQ ID NO:178), FIG. 68 (SEQ IDNO:183), FIG. 70 (SEQ ID NO:190), FIG. 73 (SEQ ID NO:196), FIG. 75 (SEQID NO:206), FIG. 77 (SEQ ID NO:211), FIG. 79 (SEQ ED NO:216), FIG. 81(SEQ ID NO:221), FIG. 83 (SEQ ID NO:226), FIG. 85 (SEQ ID NO:231), FIG.87 (SEQ ID NO:236), FIG. 89 (SEQ ID NO:245), FIG. 91 (SEQ ID NO:254),FIG. 93 (SEQ ED NO:259), FIG. 95 (SEQ ID NO:264), FIG. 98 (SEQ ID NO :270), FIG. 109 (SEQ ID NO:284), FIG. 118 (SEQ ID NO:296), FIG. 120 (SEQID NO:301), FIG. 122 (SEQ ID NO:303), FIG. 125 (SEQ ID NO:309), FIG. 129(SEQ ID NO:322), FIG. 132 (SEQ ID NO:330), FIG. 136 (SEQ ID NO:337),FIG. 139 (SEQ ID NO:346), FIG. 142 (SEQ ID NO:352), FIG. 145 (SEQ IDNO:358), FIG. 147 (SEQ ID NO:363), FIG. 149 (SEQ ID NO:370), FIG. 151(SEQ ID NO:375), FIG. 153 (SEQ ID NO:380), FIG. 155 (SEQ ID NO:385),FIG. 157 (SEQ ID NO:390), FIG. 159 (SEQ ID NO:395), FIG. 161 (SEQ IDNO:400), FIG. 163 (SEQ ID NO:405), FIG. 165 (SEQ ID NO:410), FIG. 167(SEQ ID NO:415), FIG. 169 (SEQ ID NO:420), FIG. 171 (SEQ ID NO:425),FIG. 173 (SEQ ID NO:430), FIG. 177 (SEQ ID NO:437), FIG. 179 (SEQ IDNO:442), FIG. 181 (SEQ ID NO:447), FIG. 183 (SEQ ID NO:452), FIG. 185(SEQ ID NO:454), FIG. 187 (SEQ ID NO:456), FIG. 190 (SEQ ID NO:459),FIG. 192 (SEQ ID NO:464), FIG. 194 (SEQ ID NO:466), FIG. 196 (SEQ IDNO:468), FIG. 198 (SEQ ID NO:470), FIG. 200 (SEQ ID NO:472), FIG. 202(SEQ ID NO:477), FIG. 204 (SEQ ID NO:483), FIG. 207 (SEQ ID NO:488),FIG. 209 (SEQ ID NO:496), FIG. 211 (SEQ ID NO:498), FIG. 213 (SEQ IDNO:506), FIG. 215 (SEQ ID NO:508), FIG. 217 (SEQ ID NO:510), FIG. 219(SEQ ID NO:515), FIG. 222 (SEQ ID NO:523), FIG. 225 (SEQ ID NO:526),FIG. 230 (SEQ ID NO:612), FIG. 232 (SEQ ID NO:614), FIG. 234 (SEQ IDNO:616) and FIG. 236 (SEQ ID NO:618).
 13. An isolated polypeptidescoring at least 80% positives when compared to an amino acid sequenceselected from the group consisting of the amino acid sequence shown inFIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:7), FIG. 9 (SEQ ID NO:19), FIG.11 (SEQ ID NO:28), FIG. 15 (SEQ ID NO:36), FIG. 20 (SEQ ID NO:45), FIG.22 (SEQ ID NO:52), FIG. 24 (SEQ ID NO:59), FIG. 26 (SEQ ID NO:64), FIG.28 (SEQ ID NO:69), FIG. 30 (SEQ ID NO:74), FIG. 33 (SEQ ID NO:85), FIG.35 (SEQ ID NO:90), FIG. 37 (SEQ ID NO:97), FIG. 39 (SEQ ID NO:102), FIG.41 (SEQ ID NO:109), FIG. 43 (SEQ ID NO:114), FIG. 45 (SEQ ID NO:119),FIG. 47 (SEQ ID NO:124), FIG. 49 (SEQ ID NO:132), FIG. 51 (SEQ IDNO:137), FIG. 53 (SEQ ID NO:145), FIG. 55 (SEQ ID NO:150), FIG. 59 (SEQID NO:157), FIG. 61 (SEQ ID NO:162), FIG. 63 (SEQ ID NO:169), FIG. 66(SEQ ID NO:178), FIG. 68 (SEQ ID NO:183), FIG. 70 (SEQ ID NO:190), FIG.73 (SEQ ID NO:196), FIG. 75 (SEQ ID NO:206), FIG. 77 (SEQ ID NO:211),FIG. 79 (SEQ ID NO:216), FIG. 81 (SEQ ID NO:221), FIG. 83 (SEQ IDNO:226), FIG. 85 (SEQ ID NO:231), FIG. 87 (SEQ ID NO:236), FIG. 89 (SEQID NO:245), FIG. 91 (SEQ ID NO:254), FIG. 93 (SEQ ID NO:259), FIG. 95(SEQ ID NO:264), FIG. 98 (SEQ ID NO:270), FIG. 109 (SEQ ID NO:284), FIG.118 (SEQ ID NO:296), FIG. 120 (SEQ ID NO:301), FIG. 122 (SEQ ID NO:303),FIG. 125 (SEQ ID NO:309), FIG. 129 (SEQ ID NO:322), FIG. 132 (SEQ IDNO:330), FIG. 136 (SEQ ID NO:337), FIG. 139 (SEQ ID NO:346), FIG. 142(SEQ ID NO:352), FIG. 145 (SEQ ID NO:358), FIG. 147 (SEQ ID NO:363),FIG. 149 (SEQ ID NO:370), FIG. 151 (SEQ ID NO:375), FIG. 153 (SEQ IDNO:380), FIG. 155 (SEQ ID NO:385), FIG. 157 (SEQ ID NO:390), FIG. 159(SEQ ID NO:395), FIG. 161 (SEQ ID NO:400), FIG. 163 (SEQ ID NO:405),FIG. 165 (SEQ ID NO:410), FIG. 167 (SEQ ID NO:415), FIG. 169 (SEQ IDNO:420), FIG. 171 (SEQ ID NO:425), FIG. 173 (SEQ ID NO:430), FIG. 177(SEQ ID NO:437), FIG. 179 (SEQ ID NO:442), FIG. 181 (SEQ ID NO:447),FIG. 183 (SEQ ID NO:452), FIG. 185 (SEQ ID NO:454), FIG. 187 (SEQ IDNO:456), FIG. 190 (SEQ ID NO:459), FIG. 192 (SEQ ID NO:464), FIG. 194(SEQ ID NO:466), FIG. 196 (SEQ ID NO:468), FIG. 198 (SEQ ID NO:470),FIG. 200 (SEQ ID NO:472), FIG. 202 (SEQ ID NO:477), FIG. 204 (SEQ IDNO:483), FIG. 207 (SEQ ID NO:488), FIG. 209 (SEQ ID NO:496), FIG. 211(SEQ ID NO:498), FIG. 213 (SEQ ID NO:506), FIG. 215 (SEQ ID NO:508),FIG. 217 (SEQ ID NO:510), FIG. 219 (SEQ ID NO:515), FIG. 222 (SEQ IDNO:523), FIG. 225 (SEQ ID NO:526), FIG. 230 (SEQ ID NO:612), FIG. 232(SEQ ID NO:614), FIG. 234 (SEQ ID NO:616) and FIG. 236 (SEQ ID NO:618).14. An isolated polypeptide having at least 80% amino acid sequenceidentity to an amino acid sequence encoded by the full-length codingsequence of the DNA deposited under ATCC accession number ATCC 209791,ATCC 209786, ATCC 209788, ATCC 209787, ATCC 209789, ATCC 209617, ATCC209620, ATCC 209616, ATCC 209679, ATCC 209654, ATCC 209655, ATCC 209656,ATCC 209721, ATCC 209717, ATCC 209716, ATCC 209722, ATCC 209668, ATCC209670, ATCC 209718, ATCC 209784, ATCC 209703, ATCC 209808, ATCC 209810,ATCC 209699, ATCC 209811, ATCC 209813, ATCC 209705, ATCC 209806, ATCC209809, ATCC 209805, ATCC 209812, ATCC 209844, ATCC 209847, ATCC 209845,ATCC 209843, ATCC 209846, ATCC 209750, ATCC 209848, ATCC 209851, ATCC209754, ATCC 209747, ATCC 209861, ATCC 209862, ATCC 209867, ATCC 209879,ATCC 209868, ATCC 209869, ATCC 209775, ATCC 209772, ATCC 209774, ATCC209777, ATCC 209905, ATCC 209855, ATCC 209910, ATCC 209424, ATCC 209720,ATCC 209714, ATCC 209785, ATCC 209911, ATCC 209669, ATCC 209704, ATCC209702, ATCC 209701, ATCC 209700, ATCC 209814, ATCC 209715, ATCC 209807,ATCC 209753, ATCC 209749, ATCC 209748, ATCC 209842, ATCC 209849, ATCC209880, ATCC 209864, ATCC 209882, ATCC 209883, ATCC 209865, ATCC 209866,ATCC 209857, ATCC 209870, ATCC 209859, ATCC 209653, ATCC 209389, ATCC209386, ATCC 203242, ATCC 203243, ATCC 209783, ATCC 209487, ATCC 209680,240-PTA or ATCC
 209773. 15. A chimeric molecule comprising a polypeptideaccording to any one of claims 12 to 14 fused to a heterologous aminoacid sequence.
 16. The chimeric molecule of claim 15, wherein saidheterologous amino acid sequence is an epitope tag sequence.
 17. Thechimeric molecule of claim 15, wherein said heterologous amino acidsequence is a Fc region of an immunoglobulin.
 18. An antibody whichspecifically binds to a polypeptide according to any one of claims 12 to14.
 19. The antibody of claim 18, wherein said antibody is a monoclonalantibody, a humanized antibody or a single-chain antibody.
 20. Isolatednucleic acid having at least 80% nucleic acid sequence identity to: (a)a nucleotide sequence encoding the polypeptide shown in FIG. 2 (SEQ IDNO:2), FIG. 4 (SEQ ID NO:7), FIG. 9 (SEQ ID NO:19), FIG. 11 (SEQ IDNO:28), FIG. 15 (SEQ ID NO:36), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ IDNO:52), FIG. 24 (SEQ ID NO:59), FIG. 26 (SEQ ID NO:64), FIG. 28 (SEQ IDNO:69), FIG. 30 (SEQ ID NO:74), FIG. 33 (SEQ ID NO:85), FIG. 35 (SEQ IDNO:90), FIG. 37 (SEQ ID NO:97), FIG. 39 (SEQ ID NO:102), FIG. 41 (SEQ IDNO:109), FIG. 43 (SEQ ID NO:114), FIG. 45 (SEQ ID NO:119), FIG. 47 (SEQID NO:124), FIG. 49 (SEQ ID NO:132), FIG. 51 (SEQ ID NO:137), FIG. 53(SEQ ID NO:145), FIG. 55 (SEQ ID NO:150), FIG. 59 (SEQ ID NO:157), FIG.61 (SEQ ID NO:162), FIG. 63 (SEQ ID NO:169), FIG. 66 (SEQ ID NO:178),FIG. 68 (SEQ ID NO:183), FIG. 70 (SEQ ID NO:90), FIG. 73 (SEQ IDNO:196), FIG. 75 (SEQ ID NO:206), FIG. 77 (SEQ ID NO:211), FIG. 79 (SEQID NO:216), FIG. 81 (SEQ ID NO:221), FIG. 83 (SEQ ID NO:226), FIG. 85(SEQ ID NO:231), FIG. 87 (SEQ ID NO:236), FIG. 89 (SEQ ID NO:245), FIG.91 (SEQ ID NO:254), FIG. 93 (SEQ ID NO:259), FIG. 95 (SEQ ID NO:264),FIG. 98 (SEQ ID NO:270), FIG. 109 (SEQ ID NO:284), FIG. 118 (SEQ IDNO:296), FIG. 120 (SEQ ID NO:301), FIG. 122 (SEQ ID NO:303), FIG. 125(SEQ ID NO:309), FIG. 129 (SEQ ID NO:322), FIG. 132 (SEQ ID NO:330),FIG. 136 (SEQ ID NO:337), FIG. 139 (SEQ ID NO:346), FIG. 142 (SEQ IDNO:352), FIG. 145 (SEQ ID NO:358), FIG. 147 (SEQ ID NO:363), FIG. 149(SEQ ID NO:370), FIG. 151 (SEQ ID NO:375), FIG. 153 (SEQ ID NO:380),FIG. 155 (SEQ ID NO:385), FIG. 157 (SEQ ID NO:390), FIG. 159 (SEQ IDNO:395), FIG. 161 (SEQ ID NO:400), FIG. 163 (SEQ ID NO:405), FIG. 165(SEQ ID NO:410), FIG. 167 (SEQ ID NO:415), FIG. 169 (SEQ ID NO:420),FIG. 171 (SEQ ID NO:425), FIG. 173 (SEQ ID NO:430), FIG. 177 (SEQ IDNO:437), FIG. 179 (SEQ ID NO:442), FIG. 181 (SEQ ID NO:447), FIG. 183(SEQ ID NO:452), FIG. 185 (SEQ ID NO:454), FIG. 187 (SEQ ID NO:456),FIG. 190 (SEQ ID NO:459), FIG. 192 (SEQ ID NO:464), FIG. 194 (SEQ IDNO:466), FIG. 196 (SEQ ID NO:468), FIG. 198 (SEQ ID NO:470), FIG. 200(SEQ ID NO:472), FIG. 202 (SEQ ID NO:477), FIG. 204 (SEQ ID NO:483),FIG. 207 (SEQ ID NO:488), FIG. 209 (SEQ ID NO:496), FIG. 211 (SEQ IDNO:498), FIG. 213 (SEQ ID NO:506), FIG. 215 (SEQ ID NO:508), FIG. 217(SEQ ID NO:510), FIG. 219 (SEQ ID NO:515), FIG. 222 (SEQ ID NO:523),FIG. 225 (SEQ ID NO:526), FIG. 230 (SEQ ID NO:612), FIG. 232 (SEQ IDNO:614), FIG. 234 (SEQ ID NO:616) or FIG. 236 (SEQ ID NO:618), lackingits associated signal peptide; (b) a nucleotide sequence encoding anextracellular domain of the polypeptide shown in FIG. 2 (SEQ ID NO:2),FIG. 4 (SEQ ID NO:7), FIG. 9 (SEQ ID NO:19), FIG. 11 (SEQ ID NO:28),FIG. 15 (SEQ ID NO:36), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:52),FIG. 24 (SEQ ID NO:59), FIG. 26 (SEQ ID NO:64), FIG. 28 (SEQ ID NO:69),FIG. 30 (SEQ ID NO:74), FIG. 33 (SEQ ID NO:85), FIG. 35 (SEQ ID NO:90),FIG. 37 (SEQ ID NO:97), FIG. 39 (SEQ ID NO:102), FIG. 41 (SEQ IDNO:109), FIG. 43 (SEQ ID NO:114), FIG. 45 (SEQ ID NO:119), FIG. 47 (SEQID NO:124), FIG. 49 (SEQ ID NO:132), FIG. 51 (SEQ ID NO:137), FIG. 53(SEQ ID NO:145), FIG. 55 (SEQ ID NO:150), FIG. 59 (SEQ ID NO:157), FIG.61 (SEQ ID NO:162), FIG. 63 (SEQ ID NO:169), FIG. 66 (SEQ ID NO:178),FIG. 68 (SEQ ID NO:183), FIG. 70 (SEQ ID NO:190), FIG. 73 (SEQ IDNO:196), FIG. 75 (SEQ ID NO:206), FIG. 77 (SEQ ID NO:211), FIG. 79 (SEQID NO:216), FIG. 81 (SEQ ID NO:221), FIG. 83 (SEQ ID NO:226), FIG. 85(SEQ ID NO:231), FIG. 87 (SEQ ID NO:236), FIG. 89 (SEQ ID NO:245), FIG.91 (SEQ ID NO:254), FIG. 93 (SEQ ID NO:259), FIG. 95 (SEQ ID NO:264),FIG. 98 (SEQ ID NO:270), FIG. 109 (SEQ ID NO:284), FIG. 118 (SEQ IDNO:296), FIG. 120 (SEQ ID NO:301), FIG. 122 (SEQ ID NO:303), FIG. 125(SEQ ID NO:309), FIG. 129 (SEQ ID NO:322), FIG. 132 (SEQ ID NO:330),FIG. 136 (SEQ ID NO:337), FIG. 139 (SEQ ID NO:346), FIG. 142 (SEQ IDNO:352), FIG. 145 (SEQ ID NO:358), FIG. 147 (SEQ ID NO:363), FIG. 149(SEQ ID NO:370), FIG. 151 (SEQ ID NO:375), FIG. 153 (SEQ ID NO:380),FIG. 155 (SEQ ID NO:385), FIG. 157 (SEQ ID NO:390), FIG. 159 (SEQ IDNO:395), FIG. 161 (SEQ ID NO:400), FIG. 163 (SEQ ID NO:405), FIG. 165(SEQ ID NO:410), FIG. 167 (SEQ ID NO:415), FIG. 169 (SEQ ID NO:420),FIG. 171 (SEQ ID NO:425), FIG. 173 (SEQ ID NO:430), FIG. 177 (SEQ IDNO:437), FIG. 179 (SEQ ID NO:442), FIG. 181 (SEQ ID NO:447), FIG. 183(SEQ ID NO:452), FIG. 185 (SEQ ID NO:454), FIG. 187 (SEQ ID NO:456),FIG. 190 (SEQ ID NO:459), FIG. 192 (SEQ ID NO:464), FIG. 194 (SEQ IDNO:466), FIG. 196 (SEQ ID NO:468), FIG. 198 (SEQ ID NO:470), FIG. 200(SEQ ID NO:472), FIG. 202 (SEQ ID NO:477), FIG. 204 (SEQ ID NO:483),FIG. 207 (SEQ ID NO:488), FIG. 209 (SEQ ID NO:496), FIG. 211 (SEQ IDNO:498), FIG. 213 (SEQ ID NO:506), FIG. 215 (SEQ ID NO:508), FIG. 217(SEQ ID NO:510), FIG. 219 (SEQ ID NO:515), FIG. 222 (SEQ ID NO:523),FIG. 225 (SEQ ID NO:526), FIG. 230 (SEQ ID NO:612), FIG. 232 (SEQ IDNO:614), FIG. 234 (SEQ ID NO:616) or FIG. 236 (SEQ ID NO:618), with itsassociated signal peptide; or (c) a nucleotide sequence encoding anextracellular domain of the polypeptide shown in FIG. 2 (SEQ ID NO:2),FIG. 4 (SEQ ID NO:7), FIG. 9 (SEQ ID NO:19), FIG. 11 (SEQ ID NO:28),FIG. 15 (SEQ ID NO:36), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:52),FIG. 24 (SEQ ID NO:59), FIG. 26 (SEQ ID NO:64), FIG. 28 (SEQ ID NO:69),FIG. 30 (SEQ ID NO:74), FIG. 33 (SEQ ID NO:85), FIG. 35 (SEQ ID NO:90),FIG. 37 (SEQ ID NO:97), FIG. 39 (SEQ ID NO:102), FIG. 41 (SEQ IDNO:109), FIG. 43 (SEQ ID NO:114), FIG. 45 (SEQ ID NO:119), FIG. 47 (SEQID NO:124), FIG. 49 (SEQ ID NO:132), FIG. 51 (SEQ ID NO:137), FIG. 53(SEQ ID NO:145), FIG. 55 (SEQ ID NO:150), FIG. 59 (SEQ ID NO:157), FIG.61 (SEQ ID NO:162), FIG. 63 (SEQ ID NO:169), FIG. 66 (SEQ ID NO:178),FIG. 68 (SEQ ID NO:183), FIG. 70 (SEQ ID NO:190), FIG. 73 (SEQ IDNO:196), FIG. 75 (SEQ ID NO:206), FIG. 77 (SEQ ID NO:211), FIG. 79 (SEQID NO:216), FIG. 81 (SEQ ID NO:221), FIG. 83 (SEQ ID NO:226), FIG. 85(SEQ ID NO:231), FIG. 87 (SEQ ID NO:236), FIG. 89 (SEQ ID NO:245), FIG.91 (SEQ ID NO:254), FIG. 93 (SEQ ID NO:259), FIG. 95 (SEQ ID NO:264),FIG. 98 (SEQ ID NO:270), FIG. 109 (SEQ ID NO:284), FIG. 118 (SEQ IDNO:296), FIG. 120 (SEQ ID NO:301), FIG. 122 (SEQ ID NO:303), FIG. 125(SEQ ID NO:309), FIG. 129 (SEQ ID NO:322), FIG. 132 (SEQ ID NO:330),FIG. 136 (SEQ ID NO:337), FIG. 139 (SEQ ID NO:346), FIG. 142 (SEQ IDNO:352), FIG. 145 (SEQ ID NO:358), FIG. 147 (SEQ ID NO:363), FIG. 149(SEQ ID NO:370), FIG. 151 (SEQ ID NO:375), FIG. 153 (SEQ ID NO:380),FIG. 155 (SEQ ID NO:385), FIG. 157 (SEQ ID NO:390), FIG. 159 (SEQ IDNO:395), FIG. 161 (SEQ ID NO:400), FIG. 163 (SEQ ID NO:405), FIG. 165(SEQ ID NO:410), FIG. 167 (SEQ ID NO:415), FIG. 169 (SEQ ID NO:420),FIG. 171 (SEQ ID NO:425), FIG. 173 (SEQ ID NO:430), FIG. 177 (SEQ IDNO:437), FIG. 179 (SEQ ID NO:442), FIG. 181 (SEQ ID NO:447), FIG. 183(SEQ ID NO:452), FIG. 185 (SEQ ID NO:454), FIG. 187 (SEQ ID NO:456),FIG. 190 (SEQ ID NO:459), FIG. 192 (SEQ ID NO:464), FIG. 194 (SEQ IDNO:466), FIG. 196 (SEQ ID NO:468), FIG. 198 (SEQ ID NO:470), FIG. 200(SEQ ID NO:472), FIG. 202 (SEQ ID NO:477), FIG. 204 (SEQ ID NO:483),FIG. 207 (SEQ ID NO:488), FIG. 209 (SEQ ID NO:496), FIG. 211 (SEQ IDNO:498), FIG. 213 (SEQ ID NO:506), FIG. 215 (SEQ ID NO:508), FIG. 217(SEQ ID NO:510), FIG. 219 (SEQ ID NO:515), FIG. 222 (SEQ ID NO:523),FIG. 225 (SEQ ID NO:526), FIG. 230 (SEQ ID NO:612), FIG. 232 (SEQ IDNO:614), FIG. 234 (SEQ ID NO:616) or FIG. 236 (SEQ ID NO:618), lackingits associated signal peptide.
 21. An isolated polypeptide having atleast 80% amino acid sequence identity to: (a) the polypeptide shown inFIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:7), FIG. 9 (SEQ ID NO:19), FIG.11 (SEQ ID NO:28), FIG. 15 (SEQ ID NO:36), FIG. 20 (SEQ ID NO:45), FIG.22 (SEQ ID NO:52), FIG. 24 (SEQ ID NO:59), FIG. 26 (SEQ ID NO:64), FIG.28 (SEQ ID NO:69), FIG. 30 (SEQ ID NO:74), FIG. 33 (SEQ ID NO:85), FIG.35 (SEQ ID NO:90), FIG. 37 (SEQ ID NO:97), FIG. 39 (SEQ ID NO:102), FIG.41 (SEQ ID NO:109), FIG. 43 (SEQ ID NO:114), FIG. 45 (SEQ ID NO:119),FIG. 47 (SEQ ID NO:124), FIG. 49 (SEQ ID NO:132), FIG. 51 (SEQ IDNO:137), FIG. 53 (SEQ ID NO:145), FIG. 55 (SEQ ID NO:150), FIG. 59 (SEQID NO:157), FIG. 61 (SEQ ID NO:162), FIG. 63 (SEQ ID NO:169), FIG. 66(SEQ ID NO:178), FIG. 68 (SEQ ID NO:183), FIG. 70 (SEQ ID NO:190), FIG.73 (SEQ ID NO:196), FIG. 75 (SEQ ID NO:206), FIG. 77 (SEQ ID NO:211),FIG. 79 (SEQ ID NO:216), FIG. 81 (SEQ ID NO:221), FIG. 83 (SEQ IDNO:226), FIG. 85 (SEQ ID NO:231), FIG. 87 (SEQ ID NO:236), FIG. 89 (SEQID NO:245), FIG. 91 (SEQ ID NO:254), FIG. 93 (SEQ ID NO:259), FIG. 95(SEQ ID NO:264), FIG. 98 (SEQ ID NO:270), FIG. 109 (SEQ ID NO:284), FIG.118 (SEQ ID NO:296), FIG. 120 (SEQ ID NO:301), FIG. 122 (SEQ ID NO:303),FIG. 125 (SEQ ID NO:309), FIG. 129 (SEQ ID NO:322), FIG. 132 (SEQ IDNO:330), FIG. 136 (SEQ ID NO:337), FIG. 139 (SEQ ID NO:346), FIG. 142(SEQ ID NO:352), FIG. 145 (SEQ ID NO:358), FIG. 147 (SEQ ID NO:363),FIG. 149 (SEQ ID NO:370), FIG. 151 (SEQ ID NO:375), FIG. 153 (SEQ IDNO:380), FIG. 155 (SEQ ID NO:385), FIG. 157 (SEQ ID NO:390), FIG. 159(SEQ ID NO:395), FIG. 161 (SEQ ID NO:400), FIG. 163 (SEQ ID NO:405),FIG. 165 (SEQ ID NO:410), FIG. 167 (SEQ ID NO:415), FIG. 169 (SEQ IDNO:420), FIG. 171 (SEQ ID NO:425), FIG. 173 (SEQ ID NO:430), FIG. 177(SEQ ID NO:437), FIG. 179 (SEQ ID NO:442), FIG. 181 (SEQ ID NO:447),FIG. 183 (SEQ ID NO:452), FIG. 185 (SEQ ID NO:454), FIG. 187 (SEQ IDNO:456), FIG. 190 (SEQ ID NO:459), FIG. 192 (SEQ ID NO:464), FIG. 194(SEQ ID NO:466), FIG. 196 (SEQ ID NO:468), FIG. 198 (SEQ ID NO:470),FIG. 200 (SEQ ID NO:472), FIG. 202 (SEQ ID NO:477), FIG. 204 (SEQ IDNO:483), FIG. 207 (SEQ ID NO:488), FIG. 209 (SEQ ID NO:496), FIG. 211(SEQ ID NO:498), FIG. 213 (SEQ ID NO:506), FIG. 215 (SEQ ID NO:508),FIG. 217 (SEQ ID NO:510), FIG. 219 (SEQ ID NO:515), FIG. 222 (SEQ IDNO:523), FIG. 225 (SEQ ID NO:526), FIG. 230 (SEQ ID NO:612), FIG. 232(SEQ ID NO:614), FIG. 234 (SEQ ID NO:616) or FIG. 236 (SEQ ID NO:618),lacking its associated signal peptide; (b) an extracellular domain ofthe polypeptide shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ j ID NO:7),FIG. 9 (SEQ ID NO:19), FIG. 1 (SEQ ID NO:28), FIG. 15 (SEQ ID NO:36),FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:52), FIG. 24 (SEQ ID NO:59),FIG. 26 (SEQ ID NO:64), FIG. 28 (SEQ ID NO:69), FIG. 30 (SEQ ID NO:74),FIG. 33 (SEQ ID NO:85), FIG. 35 (SEQ ID NO:90), FIG. 37 (SEQ ID NO:97),FIG. 39 (SEQ ID NO:102), FIG. 41 (SEQ ID NO:109), FIG. 43 (SEQ IDNO:114), FIG. 45 (SEQ ]OD NO:119), FIG. 47 (SEQ ID NO:124), FIG. 49 (SEQ]ID NO:132), FIG. 51 (SEQ ID NO:137), FIG. 53 (SEQ ID NO:145), FIG. 55(SEQ ID NO:150), FIG. 59 (SEQ ID NO:157), FIG. 61 (SEQ ID NO:162), FIG.63 (SEQ ID NO:169), FIG. 66 (SEQ ID NO:178), FIG. 68 (SEQ ID NO:183),FIG. 70 (SEQ ID NO:190), FIG. 73 (SEQ ID NO:196), FIG. 75 (SEQ IDNO:206), FIG. 77 (SEQ ID NO:211), FIG. 79 (SEQ ID NO:216), FIG. 81 (SEQID NO:221), FIG. 83 (SEQ ID NO:226), FIG. 85 (SEQ ID NO:231), FIG. 87(SEQ ID NO:236), FIG. 89 (SEQ ID NO:245), FIG. 91 (SEQ ID NO:254), FIG.93 (SEQ ID NO:259), FIG. 95 (SEQ ID NO:264), FIG. 98 (SEQ ID NO:270),FIG. 109 (SEQ ID NO:284), FIG. 118 (SEQ ID NO:296), FIG. 120 (SEQ IDNO:301), FIG. 122 (SEQ ID NO:303), FIG. 125 (SEQ ID NO:309), FIG. 129(SEQ ID NO:322), FIG. 132 (SEQ ID NO:330), FIG. 136 (SEQ ID NO:337),FIG. 139 (SEQ ID NO:346), FIG. 142 (SEQ ID NO:352), FIG. 145 (SEQ IDNO:358), FIG. 147 (SEQ ID NO:363), FIG. 149 (SEQ ID NO:370), FIG. 151(SEQ ID NO:375), FIG. 153 (SEQ ED NO:380), FIG. 155 (SEQ ID NO:385),FIG. 157 (SEQ ID NO:390), FIG. 159 (SEQ ID NO:395), FIG. 161 (SEQ IDNO:400), FIG. 163 (SEQ ID NO:405), FIG. 165 (SEQ ID NO:410), FIG. 167(SEQ ID NO: 415), FIG. 169 (SEQ ID NO:420), FIG. 171 (SEQ ID NO:425),FIG. 173 (SEQ ID NO:430), FIG. 177 (SEQ ID NO:437), FIG. 179 (SEQ IDNO:442), FIG. 181 (SEQ ID NO:447), FIG. 183 (SEQ ID NO:452), FIG. 185(SEQ ID NO:454), FIG. 187 (SEQ ID NO:456), FIG. 190 (SEQ ID NO:459),FIG. 192 (SEQ ID NO:464), FIG. 194 (SEQ ID NO:466), FIG. 196 (SEQ IDNO:468), FIG. 198 (SEQ ID NO:470), FIG. 200 (SEQ ID NO:472), FIG. 202(SEQ ID NO:477), FIG. 204 (SEQ ID NO:483), FIG. 207 (SEQ ID NO:488),FIG. 209 (SEQ ID NO:496), FIG. 211 (SEQ ID NO:498), FIG. 213 (SEQ IDNO:506), FIG. 215 (SEQ ID NO:508), FIG. 217 (SEQ ID NO:510), FIG. 219(SEQ ID NO:515), FIG. 222 (SEQ ID NO:523), FIG. 225 (SEQ ID NO:526),FIG. 230 (SEQ ID NO:612), FIG. 232 (SEQ ID NO:614), FIG. 234 (SEQ IDNO:616) or FIG. 236 (SEQ ID NO:618), with its associated signal peptide;or (c) an extracellular domain of the polypeptide shown in FIG. 2 (SEQID NO:2), FIG. 4 (SEQ ID NO:7), FIG. 9 (SEQ ID NO:19), FIG. 11 (SEQ IDNO:28), FIG. 15 (SEQ ID NO:36), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ IDNO:52), FIG. 24 (SEQ ID NO:59), FIG. 26 (SEQ ID NO:64), FIG. 28 (SEQ IDNO:69), FIG. 30 (SEQ ID NO:74), FIG. 33 (SEQ ID NO:85), FIG. 35 (SEQ IDNO:90), FIG. 37 (SEQ ID NO:97), FIG. 39 (SEQ ID NO:102), FIG. 41 (SEQ IDNO:109), FIG. 43 (SEQ ID NO:114), FIG. 45 (SEQ ID NO:119), FIG. 47 (SEQID NO:124), FIG. 49 (SEQ ID NO:132), FIG. 51 (SEQ ID NO:137), FIG. 53(SEQ ID NO:145), FIG. 55 (SEQ ID NO:150), FIG. 59 (SEQ ID NO:157), FIG.61 (SEQ ID NO:162), FIG. 63 (SEQ ID NO:169), FIG. 66 (SEQ ID NO:178),FIG. 68 (SEQ ID NO:183), FIG. 70 (SEQ ID NO:190), FIG. 73 (SEQ IDNO:196), FIG. 75 (SEQ ID NO:206), FIG. 77 (SEQ ID NO:211), FIG. 79 (SEQID NO:216), FIG. 81 (SEQ ID NO:221), FIG. 83 (SEQ ID NO:226), FIG. 85(SEQ ID NO:231), FIG. 87 (SEQ ID NO:236), FIG. 89 (SEQ ID NO:245), FIG.91 (SEQ ID NO:254), FIG. 93 (SEQ ID NO:259), FIG. 95 (SEQ ID NO:264),FIG. 98 (SEQ ID NO:270), FIG. 109 (SEQ ID NO:284), FIG. 118 (SEQ IDNO:296), FIG. 120 (SEQ ID NO:301), FIG. 122 (SEQ ID NO:303), FIG. 125(SEQ ID NO:309), FIG. 129 (SEQ ID NO:322), FIG. 132 (SEQ ID NO:330),FIG. 136 (SEQ ID NO:337), FIG. 139 (SEQ ID NO:346), FIG. 142 (SEQ IDNO:352), FIG. 145 (SEQ ID NO:358), FIG. 147 (SEQ ID NO:363), FIG. 149(SEQ ID NO:370), FIG. 151 (SEQ ID NO:375), FIG. 153 (SEQ ID NO:380),FIG. 155 (SEQ ID NO:385), FIG. 157 (SEQ ID NO:390), FIG. 159 (SEQ IDNO:395), FIG. 161 (SEQ ID NO:400), FIG. 163 (SEQ ID NO:405), FIG. 165(SEQ ID NO:410), FIG. 167 (SEQ ID NO:415), FIG. 169 (SEQ ID NO:420),FIG. 171 (SEQ ID NO:425), FIG. 173 (SEQ ID NO:430), FIG. 177 (SEQ IDNO:437), FIG. 179 (SEQ ID NO:442), FIG. 181 (SEQ ID NO:447), FIG. 183(SEQ ID NO:452), FIG. 185 (SEQ ID NO:454), FIG. 187 (SEQ ID NO:456),FIG. 190 (SEQ ID NO:459), FIG. 192 (SEQ ID NO:464), FIG. 194 (SEQ IDNO:466), FIG. 196 (SEQ ID NO:468), FIG. 198 (SEQ ID NO:470), FIG. 200(SEQ ID NO:472), FIG. 202 (SEQ ID NO:477), FIG. 204 (SEQ ID NO:483),FIG. 207 (SEQ ID NO:488), FIG. 209 (SEQ ID NO:496), FIG. 211 (SEQ IDNO:498), FIG. 213 (SEQ ID NO:506), FIG. 215 (SEQ ID NO:508), FIG. 217(SEQ ID NO:510), FIG. 219 (SEQ ID NO:515), FIG. 222 (SEQ ID NO:523),FIG. 225 (SEQ ID NO:526), FIG. 230 (SEQ ID NO:612), FIG. 232 (SEQ IDNO:614), FIG. 234 (SEQ ID NO:616) or FIG. 236 (SEQ ID NO:618), lackingits associated signal peptide.
 22. A method of detecting a PRO4993polypeptide in a sample suspected of containing a PRO4993 polypeptide,said method comprising contacting said sample with a PRO337 polypeptideand determining the formation of a PRO4993/PRO337 polypeptide conjugatein said sample, wherein the formation of said conjugate is indicative ofthe presence of a PRO4993 polypeptide in said sample.
 23. The methodaccording to claim 22, wherein said sample comprises cells suspected ofexpressing said PRO4993 polypeptide.
 24. The method according to claim22, wherein said PRO337 polypeptide is labeled with a detectable label.25. The method according to claim 22, wherein said PRO337 polypeptide isattached to a solid support.
 26. A method of detecting a PRO337polypeptide in a sample suspected of containing a PRO337 polypeptide,said method comprising contacting said sample with a PRO4993 polypeptideand determining the formation of a PRO4993/PRO337 polypeptide conjugatein said sample, wherein the formation of said conjugate is indicative ofthe presence of a PRO337 polypeptide in said sample.
 27. The methodaccording to claim 26, wherein said sample comprises cells suspected ofexpressing said PRO337 polypeptide.
 28. The method according to claim26, wherein said PRO4993 polypeptide is labeled with a detectable label.29. The method according to claim 26, wherein said PRO4993 polypeptideis attached to a solid support.
 30. A method of detecting a PRO1559polypeptide in a sample suspected of containing a PRO1559 polypeptide,said method comprising contacting said sample with a PRO725, PRO700 orPRO739 polypeptide and determining the formation of a PRO1559/PRO725,PRO700 or PRO739 polypeptide conjugate in said sample, wherein theformation of said conjugate is indicative of the presence of a PRO1559polypeptide in said sample.
 31. The method according to claim 30,wherein said sample comprises cells suspected of expressing said PRO1559polypeptide.
 32. The method according to claim 30, wherein said PRO725,PRO700 or PRO739 polypeptide is labeled with a detectable label.
 33. Themethod according to claim 30, wherein said PRO725, PRO700 or PRO739polypeptide is attached to a solid support.
 34. A method of detecting aPRO725, PRO700 or PRO739 polypeptide in a sample suspected of containinga PRO725, PRO700 or PRO739 polypeptide, said method comprisingcontacting said sample with a PRO1559 polypeptide and determining theformation of a PRO1559/PRO725, PRO700 or PRO739 polypeptide conjugate insaid sample, wherein the formation of said conjugate is indicative ofthe presence of a PRO725, PRO700 or PRO739 polypeptide in said sample.35. The method according to claim 34, wherein said sample comprisescells suspected of expressing said PRO725, PRO700 or PRO739 polypeptide.36. The method according to claim 34, wherein said PRO1559 polypeptideis labeled with a detectable label.
 37. The method according to claim34, wherein said PRO1559 polypeptide is attached to a solid support. 38.A method of linking a bioactive molecule to a cell expressing a PRO337polypeptide, said method comprising contacting said cell with a PRO4993polypeptide that is bound to said bioactive molecule and allowing saidPRO337 and PRO4993 polypeptides to bind to one another, thereby linkingsaid bioactive molecules to said cell.
 39. The method according to claim38, wherein said bioactive molecule is a toxin, a radiolabel or anantibody.
 40. The method according to claim 38, wherein said bioactivemolecule causes the death of said cell.
 41. A method of linking abioactive molecule to a cell expressing a PRO4993 polypeptide, saidmethod comprising contacting said cell with a PRO337 polypeptide that isbound to said bioactive molecule and allowing said PRO4993 and PRO337polypeptides to bind to one another, thereby linking said bioactivemolecules to said cell.
 42. The method according to claim 41, whereinsaid bioactive molecule is a toxin, a radiolabel or an antibody.
 43. Themethod according to claim 41, wherein said bioactive molecule causes thedeath of said cell.
 44. A method of linking a bioactive molecule to acell expressing a PRO1559 polypeptide, said method comprising contactingsaid cell with a PRO725, PRO700 or PRO739 polypeptide that is bound tosaid bioactive molecule and allowing said PRO1559 and PRO725, PRO700 orPRO739 polypeptides to bind to one another, thereby linking saidbioactive molecules to said cell.
 45. The method according to claim 44,wherein said bioactive molecule is a toxin, a radiolabel or an antibody.46. The method according to claim 44, wherein said bioactive moleculecauses the death of said cell.
 47. A method of linking a bioactivemolecule to a cell expressing a PRO725, PRO700 or PRO739 polypeptide,said method comprising contacting said cell with a PRO1559 polypeptidethat is bound to said bioactive molecule and allowing said PRO1559 andPRO725, PRO700 or PRO739 polypeptides to bind to one another, therebylinking said bioactive molecules to said cell.
 48. The method accordingto claim 47, wherein said bioactive molecule is a toxin, a radiolabel oran antibody.
 49. The method according to claim 47, wherein saidbioactive molecule causes the death of said cell.
 50. A method ofmodulating at least one biological activity of a cell expressing aPRO337 polypeptide, said method comprising contacting said cell with aPRO4993 polypeptide or an anti-PRO337 antibody, whereby said PRO4993polypeptide or said anti-PRO337 antibody binds to said PRO337polypeptide, thereby modulating at least one biological activity of saidcell.
 51. The method according to claim 50, wherein said cell is killed.52. A method of modulating at least one biological activity of a cellexpressing a PRO4993 polypeptide, said method comprising contacting saidcell with a PRO337 polypeptide or an anti-PRO4993 antibody, whereby saidPRO337 polypeptide or said anti-PRO4993 antibody binds to said PRO4993polypeptide, thereby modulating at least one biological activity of saidcell.
 53. The method according to claim 52, wherein said cell is killed.54. A method of modulating at least one biological activity of a cellexpressing a PRO1559 polypeptide, said method comprising contacting saidcell with a PRO725, PRO700 or PRO739 polypeptide or an anti-PRO1559antibody, whereby said PRO725, PRO700 or PRO739 polypeptide or saidanti-PRO1559 antibody binds to said PRO1559 polypeptide, therebymodulating at least one biological activity of said cell.
 55. The methodaccording to claim 54, wherein said cell is killed.
 56. A method ofmodulating at least one biological activity of a cell expressing aPRO725, PRO700 or PRO739 polypeptide, said method comprising contactingsaid cell with a PRO1559 polypeptide or an anti- PRO725, anti-PRO700 oranti-PRO739 antibody, whereby said PRO1559 polypeptide or saidanti-PRO725, anti- PRO700 or anti-PRO739 antibody binds to said PRO725,PRO700 or PRO739 polypeptide, thereby modulating at least one biologicalactivity of said cell.
 57. The method according to claim 56, whereinsaid cell is killed.